+

WO2018154140A1 - Diagnostic non invasif de stéatohépatite non alcoolique fibreuse - Google Patents

Diagnostic non invasif de stéatohépatite non alcoolique fibreuse Download PDF

Info

Publication number
WO2018154140A1
WO2018154140A1 PCT/EP2018/054809 EP2018054809W WO2018154140A1 WO 2018154140 A1 WO2018154140 A1 WO 2018154140A1 EP 2018054809 W EP2018054809 W EP 2018054809W WO 2018154140 A1 WO2018154140 A1 WO 2018154140A1
Authority
WO
WIPO (PCT)
Prior art keywords
nash
subject
ast
score
fibrotic
Prior art date
Application number
PCT/EP2018/054809
Other languages
English (en)
Inventor
Jérôme BOURSIER
Original Assignee
Université d'Angers
Centre Hospitalier Universitaire D'angers
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Université d'Angers, Centre Hospitalier Universitaire D'angers filed Critical Université d'Angers
Priority to ES18705960T priority Critical patent/ES2942418T3/es
Priority to US16/486,884 priority patent/US20200011869A1/en
Priority to EP18705960.5A priority patent/EP3586141B8/fr
Publication of WO2018154140A1 publication Critical patent/WO2018154140A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/08—Hepato-biliairy disorders other than hepatitis
    • G01N2800/085—Liver diseases, e.g. portal hypertension, fibrosis, cirrhosis, bilirubin
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/56—Staging of a disease; Further complications associated with the disease

Definitions

  • the present invention relates to the field of diagnosis in hepatology and more precisely to the diagnosis of non-alcohol ic steatohepatitis (NASH), preferably fib rat ic NASH.
  • NASH non-alcohol ic steatohepatitis
  • the invention also relates to the treatment of NASH, preferably fibrotic ASH.
  • Nonalcoholic steatohepatitis is an aggressive form of nonalcoholic fatty liver disease (NAFLD). NASH has the inherent propensity to progress towards liver fibrosis which can produce progressive, irreversible liver scarring and lead to cirrhosis or towards hepatocellular carcinoma (liver cancer).
  • NAFLD is characterized by the presence of hepatic steatosis, i.e., the accumulation of lipids within hepatocytes, said steatosis developing in the absence of significant alcohol consumption, use of steatogenic medication or hereditary disorders.
  • hepatic steatosis i.e., the accumulation of lipids within hepatocytes, said steatosis developing in the absence of significant alcohol consumption, use of steatogenic medication or hereditary disorders.
  • inflammation with hepatocyte injury ballooning
  • liver fibrosis Chosani N et al., Gastroenterology 2012; 142: 1592-609
  • NAFLD and NASH are general ly diagnosed through the histological examination by an expert liver pathologist of a liver sample obtained by a biopsy.
  • the NASH-CRN scoring system is commonly used by pathologists to semi-quantitatively assess the presence of steatosis, lobular inflammation, ballooning and fibrosis (Kleiner DE et al., Hepatology 2005;41 : 1313-21).
  • NAS NAFLD Activity Score
  • pathologists can allocate a score ranging from 0 to 8, corresponding to the sum. of the scores for steatosis, lobular inflammation and ballooning (Brunt EM et al., Hepatology 201 1 ;53 :810-20).
  • patients with a NAS > 4 are considered to suffer from active NASH, defined by the presence of active liver injuries that are potentially reversible in the short term.
  • active NASH defined by the presence of active liver injuries that are potentially reversible in the short term.
  • patients w ith active NASH particularly stand to benefit from therapeutic treatment since their liver injuries, while significant, can still be reversed.
  • An additional criterion to identify the patients most susceptible to benefit from treatment is the presence of fibrosis.
  • Recent cl inical trials such as the REGENERATE study (NCT02548351) or the RESOLVE-IT study (NCT02704403), are conducted in patients with biopsy-proven ASH, associated to a NAS > 4 and i gn i fi ca n t ad van ced F2-3 fibrosis, thus illustrating the clinical relevance of identifying patients w ith fibrotic NASH.
  • liver biopsy is still considered the gold standard to assess the presence and/or the severity of a liver condition in a subject, it does have limitations, notably due to a poor inter- or intra -observer reproducibility and a possible sample bias linked to the small size of the sample. Furthermore, liver biopsy is an invasive medical procedure and as such remains associated with a risk of complication and a significant cost. Accordingly, in recent years, non-invasive diagnostic tests have been developed to offer an alternative to liver biopsies. For example, Angulo et al. conceived a NAFLD Fibrosis score specifically developed for patients with NAFLD ( Angulo P et al, Hepatology 2007;45 :846-54). Polyzos SA et al. developed the CHA.
  • EP2490026 describes blood markers associated with NASH. such as I L- 1 receptor antagonist, sCD40, HMGB 1 , sPLA2 group I IA, and sPLA2 activity, and methods for detecting the presence of NASH comprising measuring at least one of these markers.
  • US2013/183687 teaches measuring and combining cleaved CK-18, intact CK-18, serum adiponectin and serum resistin to predict histological NASH.
  • WO20 14/049 1 3 1 teaches a NASH-score comprising measuring and combining BMI, CK-18, and hyperglycemia.
  • WO2015/192854 describes diagnosing NASH with NASH scores including sCD163, the level of alanine transaminase (ALT) and a value related to diabetes status. How ever, none of the existing non-invasive methods have been developed with the aim to identify patients with fibrotic NASH .
  • a non-invasive method to specifical ly and accurately assess the presence of fibrotic NASH in a subject.
  • a non-invasive test would represent a convenient and practical manner to assess the presence of fibrotic NASH in a subject, to monitor patients suffering from NAFLD or NASH for the development of fibrotic NASH and to monitor treatment response in a subject treated for fibrotic NASH.
  • the present invention thus relates to a method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject comprising measuring cytokeratin 18 (CK-18), aspartate aminotransferase (AST) and Homeostasis model assessment of insulin resistance (HOMA) in a sample of said subject.
  • CK-18 cytokeratin 18
  • AST aspartate aminotransferase
  • HOMA homeostasis model assessment of insulin resistance
  • the present invention relates to a method for assessing the presence and/or the severity of fibrotic NASH in a subject, wherein said method comprises measuring cytokeratin 18
  • the method of the invention comprises measuring cytokeratin 18 (CK-18), aspartate aminotransferase (AST) and Homeostasis model assessment of insulin resistance (HOMA) in a sample of said subject.
  • the method of the invention further comprises combining the three obtained measures in a mathematical function to obtain a score value.
  • said mathematical function is a binary logistic regression, a multiple linear regression or any multivariate analysis.
  • the method of the invention additional ly comprises comparing the score value to at least one predetermined reference value.
  • the method of the invention is computerized.
  • the sample of the subject is a serum sample, a plasma sample or a blood sample.
  • the subject is an animal, preferably a human.
  • the subject has a preexisting disease or condition, preferably metabolic syndrome and/or nonalcoholic fatty l iver disease (NAFLD).
  • NAFLD nonalcoholic fatty l iver disease
  • the method of the invention is performed on a subject who has been previously diagnosed with non-alcoholic fatty liver disease (NAFLD), and comprises the additional step of first assessing the presence of metabol ic syndrome and measuring the level of aspartate aminotransferase (AST) in a sample of said subject suffering from NAFLD, before carrying out the steps as described hereinabove.
  • NAFLD nonalcoholic fatty liver disease
  • the present invention also relates to a method for monitoring the progression of fibrotic NASH in a subject, wherein said method comprises:
  • HOMA Homeostasis model assessment of insulin resistance
  • QUICK fasting glucose-to-insulin ratio
  • the personalized reference value for a subject is a score previously determined for said subject.
  • a score is determined at least twice over a period of time.
  • the present invention also relates to a method for monitoring treatment response in a subject treated for fibrotic NASH, wherein said method comprises:
  • AST AST
  • HOMA homeostasis model assessment of insulin resistance
  • QUICK fasting glucose-to-insul in ratio
  • the determination of the score after the beginning of the treatment is repeated at least twice, and preferably at regular intervals.
  • an element means one element or more than one element.
  • ROC receiver operating characteristic
  • Biomarker or “biological marker” refers to a variable that may be measured in a sample from the subject, said sample being a bodily fluid sample, such as, for example, a blood, plasma, serum, lymph, urine, cerebrospinal fluid or sweat sample, preferably a blood, plasma, serum or urine sample, more preferably a blood, plasma or serum sample.
  • a bodily fluid sample such as, for example, a blood, plasma, serum, lymph, urine, cerebrospinal fluid or sweat sample, preferably a blood, plasma, serum or urine sample, more preferably a blood, plasma or serum sample.
  • Diagnostic target refers to the main objective of a non-invasive diagnostic test.
  • a non-invasiv e diagnostic test wherein the diagnostic target is fibrotic NASI I aims at assessing the presence and/or the severity of fibrotic NASH in a subject.
  • Fibrosis in the present invention, specifically refers to liver fibrosis, a pathological lesion of the liver made of scar tissue including fibrillary proteins or glycoproteins
  • Fibrotic NASH refers to an active NASH characterized by a NAFLD Activity Score (NAS) > 4 and the presence of significant fibrosis, defined as a fibrotic stage F > 2, preferably according to the ASH-CRN staging system.
  • NAFLD Activity Score (NAS): refers to a system of scoring the histological features of nonalcoholic fatty liver disease (NAFLD). The NAS ranges from 0 to 8, and corresponds to the sum of the scores for steatosis, lobular inflammation and ballooning. The NAS scoring system is commonly used for the histological diagnosis of NASH, defined as the presence of a score > 1 for each of the three components of the NAS. In one embodiment, a NAS score > 4 defines an active NASH.
  • NAFLD lesions encompasses all the lesions, or diseases, that correspond to the definition of nonalcoholic fatty liver disease (NAFLD) and are characterized by the presence of hepatic steatosis, i.e., the accumulation of lipids within hepatocytes.
  • NAFLD lesions include NAFLD.
  • NASH-CRN scoring system (NASH Clinical Research Network scoring system) refers to a classification system devoted to NAFLD (non-alcoholic fatty liver disease) and based on a morphological description in different classes either for steatosis (conventionally referred as grading) or fibrosis (conventionally referred as staging).
  • This semi-quantitative (ordinal in statistics) system is the most recent and conventional histological classification. This system is also known as the Kleiner grading staging system.
  • NASH nonalcoholic steatohepatitis
  • Non-invasive w hen referring to a method or test according to the present invention. means that the method or test of the invention does not comprise obtaining a tissue sample from, the body of a subject. In one embodiment, a blood sample is not considered as a tissue sample.
  • Prevalence is the proportion of patients, or rate of patients, found to be affected by the diagnostic target, i.e., the disease tested, in the population of patients considered. According to the present invention, the prevalence is expressed as a percentage. In one embodiment, the prev alence of fibrotic NASH corresponds to the proportion of patients, or rate of patients, found to have fibrotic NASH in the population of patients considered.
  • Score refers to any digit value obtained by the mathematical combination of biomarkers.
  • a score is a bound digit value, obtained by a mathematical function.
  • the score may range from 0 to 1 .
  • “Sensitivity” for a non-invasive test, measures the proportion of patients with the diagnostic target that are correctly identified as such (i.e., the percentage of patients with the diagnostic target who are correctly identified by a positive non-invasive test).
  • Specificity measures the proportion of patients without the diagnostic target that are correctly identified as such (i.e., the percentage of patients without the diagnostic target who are correctly identified by a negative non-inv asiv e test).
  • Subject refers to a mammal, preferably a human.
  • Treating refers to both therapeutic treatment and prophylactic or prev entative measures, wherein the object is to prevent or slow down (lessen) the dev elopment of NASH, preferably fibrotic NASH.
  • Those in need of treatment include those al eady with NASH, preferably fibrotic NASH, as well as those susceptible to develop NASH, preferably fibrotic NASH, or those in whom ASI I, preferably fibrotic NASH, is to be prevented.
  • a subject is successfully "treated” for NASH, preferably fibrotic NASH, if, after receiving a therapeutic amount of a therapeutic agent according to the methods of the present invention, the patient shows one or more of the following:
  • rev ersal of NASH without worsening of fibrosis, said rev ersal of NASH corresponding to the absence (score of 0) of at least one of the 3 components of the NAS (i.e., steatosis, ballooning, and inflammation); and/or
  • the present invention relates to a method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject comprising measuring eytokeratin 18
  • CK-18 aspartate aminotransferase
  • AST aspartate aminotransferase
  • HOMA Homeostasis model assessment of insulin resistance
  • the method of the invention is more accurate, both for the diagnosis of NASH and fibrotic NASH, than the tests of the prior art, such as, for example, more accurate than the NAFLD Fibrosis score (Angulo P et al., Hepatology 2007;45 :846-54) and more accurate than the NASH-scorc of WO2014 049131.
  • the Applicant has su prisingly shown that the method of the invention is more accurate than several combinations of markers described in WO2014/049131 and than the CHA index as described in Poiyzos SA et al. Ann Hepatol. 2013 Sep-Oct; 12(5):749-57.
  • the method of the invention is a non-invasive method. In one embodiment, the method of the invention is an in vitro diagnosis method.
  • NASH Nonalcoholic Fatty Liver Disease
  • NAFLD is characterized by the presence of hepatic steatosis, said steatosis developing in the absence of significant alcohol consumption, use of steatogenic medication or hereditary disorders.
  • NAFLD nonalcoholic steatohepatitis
  • NASH nonalcoholic steatohepatitis
  • AASLD nonalcoholic steatohepatitis
  • NAS ⁇ 3 patients with a NAFLD Activity Score lower than 3 (NAS ⁇ 3) arc considered to suffer from inactive NASH.
  • NAS > 4 patients with a NAFLD Activ ity Score greater than or equal to 4 (NAS > 4 ) are considered to suffer from active NASH, defined by the presence of active l iver injuries that are potential ly reversible in the short term.
  • NAS > 4 NAFLD Activity Score greater than or equal to 4
  • F > 2 fibrosis stage greater than or equal to 2
  • the method of the invention is for assessing the presence and/or the severity of NASH in a subject. In one embodiment, the method of the invention is for assessing the presence and/or the severity of active NASH in a subject. In another embodiment, the method of the invention is for assessing the presence and/or the severity of inactive NASH in a subject. In one embodiment, the method of the invention is for assessing the presence and/or the severity of fibrotic NASH in a subject.
  • the method of the invention is for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject, wherein said subject has been previously diagnosed with NAFLD.
  • the method of the invention consists in measuring cytokeratin 18 (CK-18), aspartate aminotransferase (AST) and Homeostasis model assessment of insulin resistance (HOMA ) in a sample of said subject .
  • the method of the invention does not comprise measuring any additional biomarkers.
  • the method of the invention does not comprise measuring BMI and/or hyperglycemia.
  • the method of the invention comprises:
  • CK-18 cytokeratin 18
  • AST aspartate aminotransferase
  • HOMA insulin resistance
  • the method of the invention comprises:
  • the method of the invention comprises:
  • NASH preferably fibrotic NASH
  • predetermined values of CK-18, AST and HOMA broadly encompasses any suitable reference values which may be used as a basis for comparison with respect to the measured CK-18, AST and HOMA.
  • Predetermined values of CK-1 8, AST and HOMA can be relative to a number or value derived from population studies, including without limitation, such subjects having similar age range, subjects in the same or similar ethnic group, subjects having family histories of liver condition or disease, or subjects having family histories of NAFLD.
  • the predetermined v alues of CK-18, AST and HOMA are derived from the measure of CK-18, AST and HOMA, respectively, in one or more subjects who are substantial ly healthy.
  • a "substantial ly healthy subject " ' has not been previously diagnosed or identified as having or suffering from a liver condition or disease.
  • the predetermined values of CK- 18, AST and HOMA are derived from the measure of CK-18, AST and HOMA, respectively, in one or more subjects who are diagnosed or identified as having or suffering from NAFLD, NASH, or fib rot ic NASH.
  • Another object of the invention is a method for assessing CK-18, AST and HOMA in a subject, said method comprising:
  • Caspase-cleaved cytokeratin 18 is a marker of apoptosis.
  • CK- 1 8 is released from hepatocytcs during apoptosis and can thus be measured in a sample obtained from the subject, such as in a blood, plasma or serum sample.
  • Methods for measuring CK- 1 8 include immunoassay techniques involving the use of one or more monoclonal or polyclonal antibodies. Immunoassays techniques include, without being limited to, radioimmunoassay or RIA, ELISA assays and western blot. Antibodies recognizing CK-18 are commercially available, such as for example, from Santa Cruz Biotech or Abeam.
  • the serum level of apoptotic caspase-3 generated CK.- 1 8 fragments is measured with the M30-Apoptosense enzyme-linked immunosorbent assay kit ( PEV IVA, Bromma, Sweden).
  • the level of CK- 1 8 is expressed in IU/L.
  • Aspartate aminotransferase is an enzyme mainly present in the muscles, heart and l iver. Elevated AST activity may be a marker of liver inflammation and is commonly measured in a sample obtained from the subject for diagnosing a liver impairment or disease.
  • AST activity is measured with an analyzer, such as a Roche Hitachi 91 7 (Roche Diagnostics) or a Beckman Coulter UniCel® DxC800 Synchron Clinical.
  • an analyzer such as a Roche Hitachi 91 7 (Roche Diagnostics) or a Beckman Coulter UniCel® DxC800 Synchron Clinical.
  • the level of AST is expressed in IU/L.
  • HOMA or Homeostasis model assessment of insulin resistance is a test assessing ⁇ -ceil function and insulin resistance ( I R ) from fasting glucose and insulin or C-peptide concentrations, first described in 1985 (Matthews et al., Diabetologia 28:412-419, 1985).
  • HOMA may also be referred to as HOMA-IR (Homeostasis model assessment of insulin resistance).
  • HOMA is assessed in a sample obtained from the subject using the following formula:
  • HOMA-IR [glucose (mmol/L ) * insulin ( ⁇ ⁇ )/22.5], using fasting values.
  • HOMA is assessed in a sample obtained from the subject using the logarithmic transformation of HOMA.
  • the HOMA value is capped. In one embodiment, the HOMA is capped at a value ranging from about 5 to about 20, preferably at a value ranging from about 7 to about 1 5, and more preferably is capped at about 10.
  • HOMA is a biomarker reflecting the glucose metabol ism of the subject in whom HOMA is measured.
  • alternative biomarkers reflecting the glucose metabolism of the subject may be used. Examples of such alternative markers include, without being limited to, fasting glucose-to-insul in ratio and QUICK! QU ICK!
  • QU ICKI is derived from logarithmic-transformed fasting plasma glucose (FPG) and insul in levels.
  • the measure of HOMA is replaced by the measure of the alternative biomarker fasting glucosc-to-insulin ratio. Therefore, the present invention also relates to a method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject comprising measuring CK-18, AST and fasting glucose-to-insul in ratio in a sample of said subject.
  • the present invention also relates to a method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject comprising measuring CK-18, AST and QUICKI in a sample of said subject.
  • the method of the invention for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject comprising measuring cytokeratin 18 (CK-18), aspartate aminotransferase (AST) and one of Homeostasis model assessment of insulin resistance (HOMA), fasting glucose-to-insulin ratio or QUICKI in a sample of said subject.
  • CK-18 cytokeratin 18
  • AST aspartate aminotransferase
  • HOMA Homeostasis model assessment of insulin resistance
  • QUICKI fasting glucose-to-insulin ratio
  • QUICKI fasting glucose-to-insulin ratio
  • the method of the invention does not comprise obtaining a sample from a subject.
  • the sample of the subject is a sample previously obtained from the subject. Said sample may be conserved in adequate conditions before being used in the method of the invention.
  • the sample obtained from the subject is a body fluid sample. Examples of body fluids include, w ithout being limited to, blood, plasma, serum, lymph, urine, cerebrospinal fluid or sweat.
  • the sample obtained from the subject is a blood sample. In one embodiment, the sample obtained from the subject is a whole blood sample or a plasma sample. In one embodiment, the whole blood sample or the plasma sample obtained from the subject is processed to obtain a serum sample. Methods for obtaining a serum sample from a whole blood sample or a plasma sample are routinely used in clinical laboratories.
  • the method of the invention further comprises combining the measures of CK-18, AST and HOMA in a mathematical function to obtain a score value (or alternatively combining the measures of CK-18, AST and fasting glucose-to-insulin ratio in a mathematical function to obtain a score value, or alternatively combining the measures of CK-18, AST and QUICKI in a mathematical function to obtain a score value).
  • the invention relates to a method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject com rising:
  • the method of the invention consists in :
  • the method of the invention comprises:
  • the method of the invention comprises:
  • the method of the invention comprises comparing the score value to at least one predetermined reference value.
  • reference value broadly encompasses any suitable reference value which may be used as a basis for comparison with respect to the obtained score.
  • a reference value can be relative to a number or value derived from population studies, including without limitation, such subjects having similar age range, subjects in the same or similar ethnic group, subjects hav ing family histories of l iver condition or disease, or subjects hav ing family histories of NAFLD.
  • the reference value is constructed using algorithms and other methods of statistical and structural classification.
  • the reference value is derived from the determination of the score of the invention in one or more subjects who are substantially healthy.
  • a substantially healthy subject has not been previously diagnosed or identified as having or suffering from a l iver condition or disease.
  • the reference value is derived from the determination of the score of the invention in one or more subjects who are diagnosed or identified as having or suffering from NAFLD, NASH, or fibrotic NASH.
  • the method of the invention comprises comparing the score value to a reference value, wherein :
  • a score value inferior to the reference value indicates that NASH, preferably fibrotic NASH, is not present in the subject
  • a score value superior to the reference value indicates that ASH, preferably fibrotic NASH, is present in the subject.
  • the method of the invention comprises comparing the score value to two predetermined reference values.
  • the first predetermined reference value corresponds to the exclusion cut-off and a score value below said exclusion cut-off is indicative of the absence of NASH, preferably fibrotic NASH.
  • the second predetermined reference value corresponds to the affirmation cut-off and a score value above said affirmation cut-off is indicative of the presence of ASH, preferably fibrotic NASH.
  • the interval between the two predetermined reference values, preferably between the exclusion cut-off and the affirmation cut-off corresponds to an indeterminate zone.
  • a score value within the indeterminate zone indicates that no conclusion may be drawn regarding the presence or absence of NASH, preferably fibrotic NASH, and additional tests may be required.
  • the method of the invention comprises comparing the score value to two predetermined reference values, preferably an exclusion cut-off and an affirmation cut-off, corresponding to an 80, 85, 90 or 95% sensitiv ity and an 80, 85, 90 or 95% specificity. In one embodiment, the method of invention comprises comparing the score value to two predetermined reference values, preferably an exclusion cut-off and an affirmation cut-off, corresponding to a 90% sensitivity and a 95% speci ficity.
  • the score value ranges from 0 to 1 ; the first predetermined reference value, preferably an exclusion cut-off, ranges from about 0.05 to about 0.25, preferably from about 0. 1 to about 0.2 and more preferably from about 0. 1 2 to about 0. 1 5, such as, for example, about 0.133 or about 0. 1 34; and the second predetermined reference v alue, preferably an affirmation cut-off, ranges from about 0.45 to about 0.65, preferably from about 0.5 to about 0.6 and more preferably from about 0.52 to about 0.55 or from about 0.53 to about 0.56, such as, for example, about 0.533 or about 0.550.
  • the method of the invention is for assessing the presence of NASH, preferably fibrotic NASH, in a subject and the method comprises comparing the score v alue to two predetermined reference v alues, preferably an exclusion cut-off and an affirmation cut-off, wherein :
  • a score value equal or inferior to the first predetermined reference value, preferably an exclusion cut-off, indicates that NASH, preferably fibrotic
  • NASH is not present in the subject
  • a score v alue equal or superior to the second predetermined reference v alue, preferably an affirmation cut-off, indicates that NASH, preferably fibrotic ASH, is present in the subject
  • a score value between the first and the second predetermined reference value, preferably between an exclusion cut-off and an affirmation cut-off indicates that further testing is required to assess whether NASH, preferably fibrotic NASH, is present in the subject.
  • the method of the invention is for assessing the presence of fibrotic NASH in a subject and the method comprises comparing the score value to two predetermined reference values, preferably an exclusion cut-off and an affirmation cutoff, corresponding to a 90% sensitivity and a 95% specificity, wherein the score value ranges from 0 to 1 , the first predetermined reference value, preferably an exclusion cutoff, ranges from about 0.05 to about 0.25 and the second predetermined reference value, preferably an affirmation cut-off, ranges from about 0.45 to about 0.65, and wherein :
  • a score value equal or inferior to the first predetermined reference value, preferably an exclusion cut-off. indicates that fibrotic ASH is not present in the subject
  • a score value between the first and the second predetermined reference values indicates that further testing is required to assess whether fibrotic NASH is present in the subject.
  • the mathematical function of the invention is a binary logistic regression, a multiple l inear regression or any multivariate analysis, preferably a binary logistic regression.
  • the score value is obtained by combining the measures of CK-18, AST and HOMA in a regression formula establ ished using multivariate analysis.
  • said regression is a binary logistic regression.
  • said formula is expressed as:
  • Another object of the invention is a score, obtained by measuring CK-18, AST and HOMA in a sample of a subject and combining the measures of CK-18, AST and HOMA in a mathematical function.
  • the score of the invention is obtained by measuring CK-18, AST and fasting glucose-to-insulin ratio in a sample of a subject and combining the measures of CK-18, AST and fasting glucosc-to-insulin ratio in a mathematical function.
  • the score of the invention is obtained by measuring CK- 18, AST and QUICK! in a sample of a subject and combining the measures of CK- 18 , AST and QUICK! in a mathematical function.
  • the score of the invention is obtained by measuring cytokeratin 18 (CK-18), aspartate aminotransferase (AST) and one of Homeostasis model assessment of insulin resistance (HOMA), fasting glucose-to-insulin ratio or QUICKI in a sample of a subject and combining the three obtained measures in a mathematical function.
  • the subject is a male. In another embodiment, the subject is a female.
  • the subject is an adult. According to the present invention, an adult is a subject above the age of 18, 19, 20 or 21 years. In another embodiment, the subject is a child. According to the present invention, a child is a subject below 21 , 20, 19 or 18 years, such as, for example, 1 7, 1 6, 15, 14, 13, 12, 1 1 , 10 o 9 years.
  • the subject is a patient, i.e., a warm-blooded animal, more preferably a human, who which is awaiting the receipt of, or is receiving, medical care or was/is/will be the subject of a medical procedure, or is monitored for the development or progression of a disease.
  • the subject is a human patient who is treated and/or monitored for the development or progression of a liver disease or condition (preferably NAFLD, NASH or fibrotic NASH).
  • the subject has a preexisting liver disease or condition, preferably NAFLD.
  • the subject has a preexisting disease or condition, preferably metabolic syndrome.
  • metabol ic syndrome include, without being limited to, diabetes, hypertension, dyslipidemia, a b e t a I i p o p ro t e i n e m i a , glycogen storage diseases, Weber-Christian disease, Woiman disease, and lipodystrophy.
  • the subject has a preexisting NAFLD and/or a metabolic syndrome.
  • the subject is diabetic. In another embodiment, the subject is not diabetic.
  • the method of the invention comprises the additional step of first assessing the presence of metabolic syndrome and measuring the level of aspartate aminotransferase (AST) in a sample of the subject before carrying out the steps as described hereinabove.
  • the method of the invention is performed on a subject who has been previously diagnosed with non-alcohol ic fatty l iver disease (NAFLD), and comprises the additional step of first assessing the presence of metabolic syndrome and measuring the level of aspartate aminotransferase (AST) in a sample of said subject suffering from NAFLD, before carrying out the steps as described hereinabove.
  • NASH non-alcohol ic fatty l iver disease
  • another object of the present invention is a method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject, wherein said method comprises the steps of:
  • AST aspartate aminotransferase
  • NASH preferably fibrotic NASH
  • the method of the invention is a method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a patient diagnosed with non- alcoholic fatty liver disease (NAFLD), wherein said method comprises the steps of:
  • NASH preferably fibrotic NASH
  • the level of aspartate aminotransferase is compared to a predetermined value.
  • predetermined value broadly encompasses any suitable reference value which may be used as a basis for comparison with respect to the mea.su ed level of AST.
  • a predetermined value can be relative to a number or value derived from population studies, including without limitation, such subjects having similar age range, subjects in the same or similar ethnic group, subjects having family histories of liver condition or disease, or subjects hav ing family histories of NAFLD.
  • the predetermined value is derived from the measure of AST level in one or more subjects who are substantial ly healthy.
  • a "substantially healthy subject” has not been previously diagnosed or identified as having or suffering from a liver condition or disease.
  • the predetermined value is derived from the measure of AST level in one or more subjects who are diagnosed or identified as having or suffering from
  • the predetermined value of AST ranges from about 20 UI/L to about 50 UI/L, preferably from about 25 UI/L to about 45 UI/L and more preferably from about 30 UI/L to about 40 UI/L. In one embodiment, the predetermined value of AST is about 32, 33, 34. 35. 36, 37, or 38 UI/L, more preferably about 35 UI/L.
  • carrying the method of the invention to assess the presence and/or the seventy of NASH, preferably fibrotic NASH is not required for subjects, preferably NAFLD patients, who do not have metabol ic syndrome and who have a measured AST level lower than a predetermined value, preferably lower than about 32, 33, 34, 35, 36, 37. or 38 UI/L, more preferably lower than about 35 UI/L.
  • a measured AST level lower than a predetermined value preferably lower than about 32, 33, 34, 35, 36, 37. or 38 UI/L, more preferably lower than about 35 UI/L.
  • carrying the method of the invention to assess the presence and/or the seventy of NASH, preferably fibrotic NASH, is required for subjects, preferably NAFLD patient, who also have metabolic syndrome and/or a measured AST level greater than or equal to a predetermined value, preferably greater than or equal to about 32, 33. 34. 35, 36, 37, or 38 UI I more preferably greater than or equal to about 35 UI/L.
  • the presence and/or severity of NASH should preferably be assessed by the method of the invention in patients suffering from NAFLD who also have metabol ic syndrome, in patients suffering from NAFLD who also have a measured AST level greater than or equal to a predetermined value, preferably greater than or equal to about 32, 33, 34, 35, 36, 37, or 38 UI/L, more preferably greater than or equal to about 35 UI/L, and in patients suffering from NAFLD who have both a metabolic syndrome and a measured AST level greater than or equal to a predetermined value, preferably greater than or equal to about 32, 33, 34, 35, 36, 37, or 38 UI/L, more preferably greater than or equal to about 35 UI/L.
  • the method of the invention for assessing the presence and/or the severity of ASH, preferably fib rot ic NASH, in a patient diagnosed with nonalcoholic fatty liver disease (NAFLD) comprises:
  • AST aspartate aminotransferase
  • the NAFLD patient does not have a metabolic syndrome
  • the measured level of AST is lower than a predetermined value, for example lower than about 32, 33, 34, 35, 36, 37, or 38 UI/L, in particular lower than about 35 UI/L;
  • NASI I preferably fibrotic NASH
  • the NAFLD patient has a metabolic syndrome
  • the measured level of AST is greater than or equal to a predetermined value, for example greater than or equal to about 32, 33, 34, 35, 36, 37, or 38 UI/L, in particular greater than or equal to 35 UI/L; or
  • the NAFLD patient has a metabolic syndrome and a measured level of AST greater than or equal to a predetermined value, for example greater than or equal to about 32, 33, 34, 35, 36, 37, or 38 UI/L, in particular greater than or equal to about 35 UI/L; then the presence of NASH, preferably fibrotic NASH, is to be assessed by the method of the invention as described hereinabove.
  • a measured level of AST greater than or equal to a predetermined value, for example greater than or equal to about 32, 33, 34, 35, 36, 37, or 38 UI/L, in particular greater than or equal to about 35 UI/L; then the presence of NASH, preferably fibrotic NASH, is to be assessed by the method of the invention as described hereinabove.
  • the method of the invention for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a patient diagnosed w ith non- alcoholic fatty liver disease (NAFLD) comprises:
  • CK-18 cytokeratin 18
  • AST aspartate aminotransferase
  • HOMA homeostasis model assessment of insulin resistance
  • the method is computerized.
  • the inv ention relates to a co m puter- implemented, method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject.
  • the invention relates to a computer-implemented method for assessing the presence and/or the severity of NASH, preferably fibrotic NASH, in a subject comprising:
  • Another object of the present invention is thus a computer software for implementing the method of the inv ention.
  • the method of the invention is implemented with a microprocessor comprising a software configured to calculate a score value resulting from the combinat ion of the measures of CK-18, AST and HOMA.
  • the present invention also relates to a diagnostic test for implementing the method as described hereinabove.
  • one object of the present invention is a diagnostic test comprising measuring CK-18, AST and HOMA in a sample of said subject.
  • the diagnostic test of the invention comprises measuring CK- 18, AST and HOMA in a sample of said subject and comparing the measures obtained to predetermined values of CK-18, AST and HOMA as described hereinabov e.
  • the diagnostic test of the invention comprises measuring CK-18, AST and HOMA in a sample of said subject and combining the measures of CK-18, AST and HOMA in a mathematical function to obtain a score v alue as described hereinabov e.
  • the diagnostic test of the invention comprises measuring CK- 18, AST and HOMA in a sample of said subject, combining the measures of CK- 18, AST and HOMA in a mathematical function to obtain a score value and comparing the score value to a reference value, as described hereinabove.
  • the present inv ention also relates to a kit for implementing the method as described hereinabov e.
  • the kit comprises means for measuring CK-18, AST and HOMA in a sample obtained, preferabl previously obtained, from a subject.
  • the present invention also relates to a method for monitoring the progression of NASH, preferably fibrotic NASH, in a subject, comprising measuring cytokeratin 18 (CK-18), aspartate aminotransferase (AST) and Homeostasis model assessment of insul in resistance (HOMA ) in a sample of the subject at least twice, and preferably at regular intervals.
  • CK-18 cytokeratin 18
  • AST aspartate aminotransferase
  • HOMA Homeostasis model assessment of insul in resistance
  • said method comprises:
  • measuring CK-18, AST and HOMA in a sample of the subject optionally comparing the measures obtained for CK-18, AST and HOMA to predetermined values of CK-18, AST and HOMA; repeating the measures at least twice, and preferably at regular intervals.
  • the present invention also relates to a method for monitoring the progression of NASH, preferably fibrotic NASH, in a subject, wherein said method comprises:
  • the reference value is a personalized reference value.
  • the personalized reference value for a subject is the first score value obtained for said subject.
  • said method comprises:
  • determining a score by measuring in a sample of said subject CK-18, AST and HOMA and combining the measures of CK-18, AST and HOMA in a mathematical function as described hereinabove, optionally comparing the score value to a reference value as described hereinabove;
  • the method for monitoring the progression of NASH, preferably fibrotic NASH, in a subject comprises:
  • determining a score by measuring in a sample of said subject CK-18, AST and HOMA and combining the measures of CK-18, AST and HOMA in a mathematical function as described hereinabove;
  • a score is determined at least twice (such as, for example, twice, three times, four times or more) over a period of time. In one embodiment, the score is determined once a week, once a month or once a year. In one embodiment, the determination of the score is repeated every 6 months, every 9 months or every 18 months. In another embodiment, the determination of the score is repeated every year, every 2 years, every 3 years, every 4 years or every 5 years.
  • the present invention also relates to a method for treatin NASH, preferably fibrotic NASH, in a subject in need thereof, comprising identifying the subject to treat by measuring in a sample of said subject cytokeratin 18 (CK-18), aspartate aminotransferase (AST) and Homeostasis model assessment of insulin resistance (HOMA).
  • CK-18 cytokeratin 18
  • AST aspartate aminotransferase
  • HOMA Homeostasis model assessment of insulin resistance
  • the method comprises identifying the subject to treat by measuring in a sample of said subject CK-18, AST and HOMA and comparing the measures obtained for CK-18, AST and HOMA to predetermined values of CK-18, AST and HOMA.
  • the method comprises identifying the subject to treat by measuring in a sample of said subject CK-18, AST and HOMA and combining the measures of CK-18, AST and HOMA in a mathematical function to obtain a score value as described hereinabove, optionally comparing the score value to a reference value as described hereinabove.
  • the method for treating NASH, preferably fibrotic NASH, in a subject in need thereof comprises:
  • the at least one therapeutic agent is treating NASH, preferably fibrotic NASH, and/or amel iorating or alleviating the symptoms associated with NASH, preferably fibrotic NASH.
  • Another object of the invention is at least one therapeutic agent for use in the treatment of NASH, preferably fibrotic NASH in a subject, wherein the subject to be treated is identified by: measuring in a sample of said subject CK-18, AST and HOMA; and combining the measures of CK-18, AST and HOMA in a mathematical function to obtain a score value as described hereinabove, optionally comparing the score value to at least one reference value as described hereinabove.
  • the at least one therapeutic agent is selected from the group comprising: drugs regulating metabolism homeostasis (such as, for example, thiazolidine derivatives, obetichoiic acid, elafibranor); drugs regulating oxidative stress (such as, for example, vitamin E, ask- 1 inhibitor); drugs regulating inflammation (such as, for example, cenicriviroc); drugs regulating apoptosis (such as, for example, the caspase inhibitor Emricasan ); and anti-fibrotic drugs (such as, for example, pumpuzumab).
  • drugs regulating metabolism homeostasis such as, for example, thiazolidine derivatives, obetichoiic acid, elafibranor
  • drugs regulating oxidative stress such as, for example, vitamin E, ask- 1 inhibitor
  • drugs regulating inflammation such as, for example, cenicriviroc
  • drugs regulating apoptosis such as, for example, the caspase inhibitor Emricasan
  • the at least one therapeutic agent is selected from the group comprising insul in sensitizers (such as rosiglitazone, pioglitazone and MSDC-0602K); farnesoid X receptor ( FXR ) agonists (such as obetichoi ic acid (also referred to as OCA ), GS-9674, LJN452, LMB763 and EDP-305); Peroxisome Proliferator-Activated Receptor ⁇ / ⁇ (PPAR ⁇ / ⁇ ) agonists (such as elafibranor, sarogl itazar and IVA337); fibroblast growth factor 1 9 (FGF19) analogs (such as NGM282); fibroblast growth factor 21 (FGF21) analogs (such as PF-0523 1023 ); recombinant FGF2 1 (such as BMS-986036); stearoyi- coenzymc A desaturase 1 (SCD 1) inhibitors (such as BMS
  • the at least one therapeutic agent is selected from the group comprising obeticholic acid; elafibranor; cenicriviroc; ask-1 inhibitor; thiazol idine derivatives; and vitamin E.
  • the at least one therapeutic agent is selected from the group comprising thiazolidine derivatives, preferably pioglitazone or rosiglitazone; antioxidants, preferably vitamin E and/or vitamin C; obeticholic acid; elafibranor; cenicriviroc; and ask-1 inhibitor; and any combination thereof.
  • the at least one therapeutic agent is obeticholic acid or elafibranor.
  • the present invention also relates to a method for monitoring treatment response in a subject treated for NASH, preferably fibrotic NASH, wherein said method comprises measuring cytokeratin 18 (C -18), aspartate aminotransferase (AST) and Homeostasis model assessment of insulin resistance (HOMA) in a sample of the subject at least twice, and preferably at regular intervals.
  • the measure is performed at least once before the beginning of the treatment and at least once after the beginning of the treatment.
  • the method of the invention comprises:
  • the method of the invention comprises comparing the measures or the scores obtained to a reference value.
  • the term "reference value" is defined hereinabove.
  • the reference value is a personal ized reference value.
  • the personal ized reference value for a subject is the score value obtained for said subject before the beginning of treatment.
  • the method of the invention comprises: measuring CK-18, AST and HOMA in a sample of the subject before the beginning of the treatment; and
  • the method of the invention comprises:
  • the method for monitoring treatment response in a subject treated for NASH, preferably fibrotic NASH comprises:
  • the determination of the score after the beginning of the treatment is repeated at regular intervals.
  • the determination of the score is repeated every month, every 2 months, every 3 months, every 6 months or every 9 months after the beginning of the treatment.
  • the determination of the score is repeated every year, every 2 years, every 3 years, every 4 years or every 5 years after the beginning of the treatment.
  • a significant decrease in a score determined after the beginning of the treatment as mentioned hereinabove as compared to the score determined before the beginning of the treatment indicates that the treatment is effective.
  • a significant decrease in a score is a decrease of at least about 2,5%, 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%. 40%, 42.5%, 45%. 47.5% or 50% as compared to the score determined before the beginning of the treatment.
  • a significant decrease in a score is a decrease greater than about 2,5%, 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5% or 50% as compared to the score determined before the beginning of the treatment.
  • the method of the invention specifical ly allows to assess the presence of fibrotic NASH .
  • the Appl icant demonstrated that the new score of the invention (that may also be referred to as MACK-3 test), combining AST, HOMA and CK-18, preferably in a binary logistic regression, has a very good accuracy for the non-invasive diagnosis of fibrotic NASH with 93.3% of correctly classified patients, a sensitivity of 90.0%, and a specificity of 94.2%, and with AUROC at 0.846 ⁇ 0.016.
  • AUROC area under the ROC curve
  • AUROC between 0.5 and 0.7 indicates that the test has poor to fair discrimination.
  • AUROC between 0.7 and 0.8 indicates that the test has acceptable discrimination
  • AUROC betw een 0.8 and 0.9 indicates that the test has excel lent discrimination ( Forthofer RN, Lee ES, Hernandez M (2006) Biostatistics: A guide to Design, Analysis, and Discovery: Elsevier).
  • MACK-3 test of the invention has a significantly superior diagnostic accuracy compared to that of previously described tests.
  • MACK-3 is more accurate than the NALFD Fibrosis Score ( NFS ) and the NASH-score of WO2014/049131 , both for NASH and for fibrotic NASH.
  • NFS NALFD Fibrosis Score
  • NASH-score WO2014/049131
  • MACK-3 has an AUROC of 0.846 ⁇ 0.016 (see Tables 8 and 14) whereas NFS has an AUROC of 0.654 ⁇ 0.023 (see Tables 4 and 8) and NASH-score has an AUROC of 0.647 ⁇ 0.023 (see Table 14 ).
  • the MACK-3 test of the invention also displays a significantly superior diagnostic accuracy compared to that of untested combinations disclosed in WO2014 0491 1 , i.e., the combinations of BM I, CK-18 and glycemia (combination B); and CK-18, weight and insul in (combination C).
  • MACK-3 has an AUROC of 0.846 ⁇ 0.016 (see Tables 8, 14 and 15) whereas combination A has an AUROC of 0.757 ⁇ 0.021 , combination B has an AUROC of 0.784 ⁇ 0.019 and combination C has an AUROC of 0.736 ⁇ 0.022 (see Table 1 5 ).
  • AUROC 0.846 ⁇ 0.016
  • combination A has an AUROC of 0.757 ⁇ 0.021
  • combination B has an AUROC of 0.784 ⁇ 0.019
  • combination C has an AUROC of 0.736 ⁇ 0.022 (see Table 1 5 ).
  • the differences in diagnostic accuracy between MACK-3 and combinations A. B and C are significant with a p-value lower than 0.050.
  • the MACK-3 test of the invention displays a surprising and significant superiority in diagnostic accuracy when compared to several combinations of markers described in WO2014 0491 3 1 .
  • the MACK-3 test of the invention displays a significantly superior diagnostic accuracy compared to that of the CHA index as described in Poiyzos SA et al. Ann Hepatol . 2013 Sep-Oct; 12(5):749-57.
  • the overal l diagnostic accuracy of MACK-3 is about 93% of patients correctly classified whereas that of the CHA index is about 75.5% of patients correctly classified (see Table 17).
  • the MACK-3 test of the invention also displays a surprising and significant superiority in diagnostic accuracy when compared to the CHA inde as described in Poiyzos SA et al. Ann Hepatol . 201 3 Sep-Oct; 12(5):749-57.
  • Fibrotic NASH is a particularly relevant diagnostic target in the clinical practice. Indeed, among the patients suffering from NAFLD or NASH, patients with fibrotic NASH particularly stand to benefit from therapeutic treatment since their liver injuries, while significant, can still be reversed.
  • the significance of fibrotic NASH is illustrated by the fact that criteria defining fibrotic NASH, i.e., an active ASH with a NAS > 4 and a significant fibrosis (fibrosis stage F > 2, preferably according to the N ASH-CRN staging system), arc the inclusion criteria of on-going cl inical trials aiming to assess the safety and efficacy of therapeutic agents in subjects with NASH .
  • fibrotic NASH is now used in the stratification of NAFLD severity by the latest international HAS L E A S D E A S O Clinical Practice Guidelines (EASL. EASL-EASD-EASO Clinical Practice Guidelines for the management of non-alcohol ic fatty liver disease. J Hepatol 2016;64: 1388-1402).
  • the method and diagnostic test of the invention thus represent a simple and accurate solution to improve the screening and monitoring of patients for therapeutic trials in NASH . Indeed, inclusions in pha.se I I therapeutic trials in the setting of NASH are currently challenged by high rates of screening failure. Since it allows the identification of a subset of NAFLD patients with a high rate of fibrotic N ASH (see Figure 4), the MACK-3 test of the invention should considerably reduce the rate of screening failure. Furthermore, the method and diagnostic test of the invention allows the non-invasive identi fication and monitoring of the patients who are l ikely to benefit the most from the newly developed NASH therapies.
  • NAFLD now concerns 25% of the general population (Younossi ZM et al. Hepatology 2016;64:73-84). Consequently, with the upcoming arrival of new treatments, many patients will need to be tested for the presence of fibrotic NASH.
  • One of the methods of the invention (also referred to as MACK-3 algorithm ) allows to l imit extensive measurements of specialized biomarkers in patients with NAFLD (see Figure 5) and thus to limit costs related to the use of the MACK-3 test in clinical practice.
  • Figure 1 is a diagram showing the prevalence of liver lesions in the study population of 846 patients of Example 1 ( ASH : nonalcohol ic steatohepatitis; NAS: NAFLD Activity
  • Figure 2 is a combination of graphs showing the accuracy for the diagnosis of fibrotic NASH of the NAFLD fibrosis score (NFS ) and the FIB4 test, each performed with their published diagnostic cut-offs: for NFS - 1 .455 and 0.676. as described in Angulo P et al., Hepatology 2007;45 :846-54 and for FIB4 1 .30 and 2.67, as described in Shah AG et al.,
  • the accuracy of the NFS and FIB4 tests is assessed through the determination (A ) of the proportion of all fibrotic NASH patients included in the di fferent intervals of the NFS and FIB4 tests; and (B) of the prevalence of fibrotic NASH as a function of the diagnostic interval of the NFS and FIB4 tests.
  • Figure 3 is a combination of graphs showing the CK-18 level of patients (expressed in IU/L) as a function of the presence in the patients of: NASH and/or NAS > 4 (A) or NASH and/or NAS > 4 with fibrosis stage F > 2, the latter defined as either fibrosis stage F0- 1 or fibrosis stage F2-4 (B).
  • Figure 4 is a graph showing the prevalence of the NAFLD lesions (advanced fibrosis without active NASH; fibrotic NASH; active NASH with no ; mild fibrosis; and no active NASH and no advanced fibrosis) as a function of the three interv als of MACK-3 score in both the derivation and the v alidation population sets.
  • Figure 5 is a diagram il lustrating the MACK-3 algorithm for the diagnosis of fibrotic
  • Figure 6 is a graph showing the prevalence of the NAFLD lesions (advanced fibrosis w ithout active NASH; fibrotic NASH; active NASH with no mild fibrosis; and no active NASH and no advanced fibrosis) as a function of the four subgroups defined by the MACK-3 algorithm.
  • Example 1 The present invention is further il lustrated by the following examples.
  • Example 1 The present invention is further il lustrated by the following examples.
  • Example 1 Example 1:
  • the study population was obtained by pooling the data from three published studies performed in Angers (Boursier J et al., J Hepatol. 2016 Sep;65(3):570-8), Antwerp ( Francque SM et al, Clin Gastroenterol Hepatol 2012; 10: 1 162-8) and N ice (Amy R et al, Aliment Pharmacol Ther 2010;32: 13 1 5-22 ).
  • patients had to hav e biopsy- prov en NAFLD as defined by the presence of liver steatosis after exclusion of concomitant steatosis-inducing drugs (such as corticosteroids, tamoxifen, amiodarone, or methotrexate), excessiv e alcohol consumption (> 30 g day in men or > 20 g day in women ), chronic hepatitis B or C infection, and histological evidence of other concomitant chronic l iver disease. Patients were excluded if they had cirrhosis complication (hepatic encephalopathy, ascites, variceal bleeding, systemic infection, or hepatocellular carcinoma).
  • concomitant steatosis-inducing drugs such as corticosteroids, tamoxifen, amiodarone, or methotrexate
  • excessiv e alcohol consumption > 30 g day in men or > 20 g day in women
  • Angers center included outpatients from the hepatology department
  • Antwerp center included obese patients without pre-existing diabetes who presented at the obesity clinic for a problem of overweight
  • ice center included morbidly obese patients referred for a bariatric surgery.
  • Metabolic syndrome was defined as the presence of at least three of the following parameters (Albert!
  • elevated waist circumference >94 cm in men or >80 cm in women
  • elevated blood pressure systolic blood pressure > 1 30 mm Hg or diastolic blood pressure >85 mm Hg or antihypertensive drug
  • elevated giycemia > 1 00 mg/dL or anti-diabetic drug
  • elevated triglycerides > 1 50 mg/dL or lipid-lowering drug
  • low HDL cholesterol ⁇ 40 mg/dL in men or ⁇ 50 mg/dL in women or l ipid-lowering drug.
  • Liver biopsy was performed percutaneously ( 16-gauge Menghini ) in Angers, percutaneously ( 16-gauge Menghini ) or peri-operatively ( 14-gaugc Tru-Cut) in Antwerp, or during bariatric surgery (hepatic wedges) in Nice.
  • Pathological examination of liver biopsies was performed by one (Angers, Antwerp) or two (Nice) senior expert(s) specialized in hepatology who were blinded for patient data. Steatosis, lobular inflammation and ballooning were semi-quantitatively graded using the N ASH-CRN scoring system (Kleiner DE et al., Hepatology 2005:41 : 13 1 -2 1 ).
  • the NAS corresponded to the sum of the scores for steatosis, lobular inflammation and ballooning ( Brunt EM et al, Hepatology 201 1 ;53:810-20).
  • NASH was defined as the presence of a score > 1 for each of the three components of the NAS.
  • Active NASH was defined as the presence of NASH with a NAS score > 4.
  • Liver fibrosis was staged from F0 to F4 according to the NASH-CRN scoring system (Kleiner DE et al, Hepatology 2005;4 1 : 13 13-2 1 ). Significant fibrosis was defined as F > 2 and advanced fibrosis as F > 3. Final ly, "fibrotic NASH” was defined as the presence of an active ASH with significant fibrosis, i.e., NASH and NAS > 4 and fibrosis F > 2. Fibrotic ASH was the primary diagnostic target of the study.
  • Liver biopsies were digitized using an Aperio digital slide scanner (Scanscope CS System, Aperio Technologies, Vista CA 92081 , USA) image processor that provided high quality images (30,000 x 30,000 pixels) at a resolution of 0.5 Li m pixel (magnification x20). Two slides were digitized for each patient: one stained with hematoxylin and cos in and one with picrosirius red. The digitized sl ides were sent to the expert pathologist from each of the three investigating centers to score steatosis, lobular inflammation, ballooning, and fibrosis according to the N ASH-CRN scoring system.
  • Aperio digital slide scanner Scanscope CS System, Aperio Technologies, Vista CA 92081 , USA
  • HO MA was calculated as fol lows: fasting glucose (mmoi/L) * fasting insul in ( ⁇ /mL) / 22.5.
  • HO MA was capped at 10.
  • Serum level of apoptotic caspa.se- 3 generated C - 1 8 fragments was measured on frozen samples stored at -80°C in each center, using the M30-Apoptosense enzyme-l inked immunosorbent assay kit (PEVIVA, Bromma, Sweden ). All blood assays were performed in the laboratories of the three participating centers. Blood fibrosis tests demonstrably have excellent inter-laboratory reproducibility (Caies P et al, Clin Bioehcm 2008:41 : 10- 1 8 ).
  • BMI body mass index
  • AST aspartate aminotransferase
  • ALT alanine aminotransferase
  • GGT gamma glutamyl-transpeptidase
  • HbAlc hemoglobin Ale also known as glycated hemoglobin.
  • liver biopsies of 30 patients were all read by the expert pathologist from each investigating center, for a total of 90 observations.
  • the distribution of the recorded scores is shown in Table 2 below.
  • Fleiss' Kappa score was 0.77 for steatosis, 0.62 for lobular inflammation, 0.64 for ballooning, 0.58 for NASH, 0.81 for the fibrosis stage, and 0.60 for fibrotic NASH. As shown in Table 3 below, these results were very close to what has been previously observed between nine experts of the NASH-CRN.
  • FIB4 had a higher AUROC (0.732 ⁇ 0.021) than NFS (0.654 ⁇ 0.023, p ⁇ 0.001) and BARD (0.566 ⁇ 0.023, p ⁇ 0.001).
  • CK-18 was significantly correlated to steatosis, lobular inflammation, ballooning and fibrosis with significant difference between most of the adjacent grades/stages.
  • CK-18 was significantly higher in patients having NASH compared to those without (respectively: 319 ⁇ 316 vs 217 ⁇ 142 IU/L, p ⁇ 0.001).
  • CK-18 surprisingly showed a low accuracy for the diagnosis of NASH with an AUROC at 0.595 ⁇ 0.019.
  • the level of CK-18 as a function of NASH, NAS > 4 and F > 2 was then evaluated.
  • CK-18 level did not significantly differ between patients with NASH and patients without NASH.
  • the level of CK-18 was significantly higher in F > 2 patients compared to FO-1 patients ( Figure 3B). Consequently, as seen in Table 4 hereinabove, rather than a biomarker for the single diagnosis of NASH, CK-18 was more globally associated with the severity of NAFLD. CK-18 performed better in diagnosing fibrotic NASH and appeared as a candidate biomarker for this diagnostic target.
  • Table 5 CK-18 level as a function of NAFLD lesions
  • BMI body mass index
  • AST aspartate aminotransferase
  • ALT alanine aminotransferase
  • GGT gamma glutamyl-transpeptidase
  • HbAlc hemoglobin Ale also known as glycated hemoglobin.
  • b waist circumference > 94 cm in men or > 80 cm in women
  • c systolic blood pressure > 130 mm Hg and/or diastolic blood pressure > 85 mm Hg and/or antihypertensive drug
  • d glycemia 100 mg/dL or anti-diabetic drug
  • e triglycerides > 150 mg/dL or lipid- lowering drug
  • h glycemia > 126 mg/dL or anti-diabetic drug;
  • s NASH and NAS > 4 and fibrosis F > 2
  • the 846 patients were 2: 1 randomly divided into a derivation set (564 patients) and a validation set (282 patients), with patient characteristics not being significantly different in the two sets, as shown in Table 6 above.
  • a derivation set 564 patients
  • a validation set 282 patients
  • patient characteristics not being significantly different in the two sets, as shown in Table 6 above.
  • multivariate analysis identified three variables as independent predictors of fibrotic NASH: AST, HOMA and CK-18 (see Table 7 below).
  • the regression formula (binary logistic regression) of the multivariate analysis was used to derive a new blood test combining these three parameters: the MACK-3 (HOMA, AST, CK-18) resulting in a score value ranging from 0 to 1.
  • the MACK-3 thresholds corresponding to 90% sensitivity and 95% specificity for fibrotic NASH were calculated in the derivation population at, respectively, 0.134 and 0.550.
  • MACK-3 had a significantly higher AUROC for the diagnosis of fibrotic NASH (0.847 ⁇ 0.030) than BARD, NFS and FIB4 (p ⁇ 0.002, see Table 8 below).
  • NFS and FIB4 showed only moderate diagnostic accuracy for fibrotic NASH with AUROC respectively at 0.618 ⁇ 0.040 and 0.721 ⁇ 0.0.037.
  • the new MACK-3 test combining HOMA, AST, CK-18 showed a very good accuracy for the non-invasive diagnosis of fibrotic NASH with AUROC at 0.846 ⁇ 0.016.
  • MACK-3 ⁇ 0.134 48.3% of the patients had MACK-3 ⁇ 0.134 (90% sensitivity threshold), 36.0% were in the intermediate grey zone (0.135-0.549), and 15.7% had MACK-3 > 0.550 (95% specificity threshold).
  • MACK-3 ⁇ 0.134 provided 90.0% sensitivity for fibrotic NASH, and MACK-3 > 0.550 had 94.2% specificity.
  • 85.7% had fibrotic NASH or advanced F3-4 fibrosis ( Figure 4).
  • MACK-3 showed an excellent diagnostic accuracy for fibrotic NASH with 93.3%) well-classified patients as shown in Table 9 below. All the results were not significantly different between the derivation and the validation sets.
  • DA overall diagnostic accuracy (i.e., rate of well-classified patients, %); Se: sensitivity (%); Spe: specificity (%); NPV: negative predictive value (%); PPV: positive predictive value; -LR: negative likelihood ratio; +LR: positive likelihood ratio; OR: diagnostic odds ratio; LB: liver biopsy requirement (%).
  • DA overall diagnostic accuracy (i.e., rate of well-classified patients, %); Se: sensitivity (%); Spe: specificity (%); NPV: negative predictive value (%); PPV: positive predictive value; -LR: negative likelihood ratio; +LR: positive likelihood ratio; OR: diagnostic odds ratio; LB: liver biopsy requirement (%)
  • NAFLD represents a very large population in which using MACK-3 to identify the subgroup of patients who need to be treated would require extensive measurement of CK18 and HOMA-IR.
  • simple parameters available in routine practice (age, sex, BMI, MetS, AST, ALT, GGT) can be used in a first step to identify a subgroup with no need for MACK-3 calculation because of a very low prevalence of fibrotic NASH.
  • multivariate analysis including the seven simple parameters listed above identified AST and MetS (metabolic syndrome) as independent predictors of fibrotic NASH. The best cut-off for AST was calculated at 35 IU/L.
  • MACK-3 algorithm a new algorithm, referred to as the "MACK-3 algorithm” was derived, according to which MACK-3 is only used for the diagnosis of fibrotic NASH in clinical practice in patients having MetS and/or AST > 35 UI/L, as depicted in Figure 5. Patients with either MetS or ALT > 35 IU/L were stratified according to the three intervals of MACK-3. As shown in Table 12 below, the accuracy of the MACK-3 algorithm was not significantly different between the derivation and the validation sets.
  • the rates of patients included in the four subgroups of the MACK-3 algorithm were as follow: 25.6% of patients in the "no MetS and AST ⁇ 35UI/L" subgroup, 26.1% in the "MetS and/or AST > 35 IU/L with MACK-3 ⁇ 0.134" subgroup, 34.4% of patients in the "MetS and/or AST > 35 IU/L with 0.135 ⁇ MACK-3 ⁇ 0.549" subgroup, and 13.9% of patients in the "MetS and/or AST > 35 IU/L with MACK-3 > 0.550" subgroup.
  • the prevalence of fibrotic NASH in the four subgroups of the MACK-3 algorithm was, respectively: 1.0% in the "no MetS and AST ⁇ 35UI/L” subgroup, 8.7% in the "MetS and/or AST > 35 IU/L with MACK-3 ⁇ 0.134" subgroup, 33.5% in the "MetS and/or AST > 35 IU/1 with 0.135 ⁇ MACK-3 ⁇ 0.549" subgroup and 69.1% "MetS and/or AST > 35 IU/L with MACK-3 > 0.550" subgroup.
  • Figure 6 shows that among the patients included in the last subgroup (MetS and/or AST > 35 UI/L with MACK-3 > 0.550), 82.7% had fibrotic NASH or advanced F3-4 fibrosis. Only 10.7% of the patients with fibrotic NASH and 15.3% of the patients with advanced F3-4 fibrosis were missed by the MACK-3 algorithm, which respectively represented 2.5%> and 2.8%> of the whole population. Finally, the MACK-3 algorithm provided an excellent accuracy for the diagnosis of fibrotic NASH with 93.2%> well-classified patients, 89.3% sensitivity, 94.4%> specificity, 83.1%) positive predictive value, and 96.6%> negative predictive value as indicated in Table 12 below. Table 12: Accuracy of the MACK-3 algorithm for the diagnosis of fibrotic NASH
  • DA overall diagnostic accuracy (i.e., rate of well-classified patients, %>); Se: sensitivity (%>); Spe: specificity (%>); NPV: negative predictive value (%>); PPV: positive predictive value; -LR: negative likelihood ratio; +LR: positive likelihood ratio; OR: diagnostic odds ratio; LB: liver biopsy requirement (%>)
  • Example 1 The study was conducted on the population described in Example 1 , obtained by pooling the data from three published studies performed in Angers (Boursier J et al., J Hepatol. 2016 Sep;65(3):570-8), Antwerp (Francque SM et al., Clin Gastroenterol Hepatol 2012;10:1162-8) and Nice (Anty R et al., Aliment Pharmacol Ther 2010;32: 1315-22.). Their characteristics are presented hereinabove in Example 1 and detailed in Table 1.
  • the 846 patients were 2: 1 randomly divided into a derivation set (564 patients) and a validation set (282 patients), with patient characteristics not being significantly different in the two sets, as shown hereinabove in Table 6.
  • MACK-3 is the combination of the three biomarkers HOMA, AST, and CK-18 in a binary logistic regression.
  • NASH-score is described in WO2014/049131 and corresponds to the combination of the three biomarkers BMI, CK-18, and hyperglycemia. Both blood tests were performed in the derivation population, in the validation population and in the whole study population.
  • the new MACK-3 test combining HOMA, AST, CK- 18 in a binary logistic regression, showed a good accuracy for the non-invasive diagnosis of NASH with AUROC at 0.785 ⁇ 0.028 in the validation set.
  • the NASH- score combining BMI, CK-18, and hyperglycemia showed a poor accuracy with AUROC at 0.497 ⁇ 0.035 in the validation set. Similar results were obtained in the derivation set and in the whole study population.
  • MACK-3 showed a significantly greater accuracy than NASH-score for the diagnosis of NASH.
  • Table 13 AUROC of MACK-3 and NASH-score for the diagnosis of NASH
  • MACK-3 showed a very good accuracy for the non-invasive diagnosis of fibrotic NASH with AUROC at 0.847 ⁇ 0.030 in the validation set.
  • NASH-score showed a moderate accuracy with AUROC at 0.636 ⁇ 0.042 in the validation set. Similar results were obtained in the derivation set and in the whole study population.
  • MACK-3 showed a significantly greater accuracy than NASH-score for the diagnosis of fibrotic NASH.
  • Example 1 The study was conducted on the population described in Example 1 , obtained by pooling the data from three published studies performed in Angers (Boursier J et al., J Hepatol. 2016 Sep;65(3):570-8), Antwerp (Francque SM et al., Clin Gastroenterol Hepatol 2012;10:1162-8) and Nice (Anty R et al., Aliment Pharmacol Ther 2010;32: 1315-22). Their characteristics are presented hereinabove in Example 1 and detailed in Table 1. The 846 patients were 2: 1 randomly divided into a derivation set (564 patients) and a validation set (282 patients), with patient characteristics not being significantly different between the two sets, as shown hereinabove in Table 6.
  • MACK-3 is the combination of the three biomarkers HOMA, AST, and CK-18 in a binary logistic regression.
  • the combination A corresponds to the combination of CK-18, BMI, and HOMA.
  • the combinations B and C are described in WO2014/049131 and corresponds to the combinations of:
  • combination B BMI, CK-18 and glycemia
  • combination C CK-18, weight and insulin.
  • MACK-3 showed a very good accuracy for the non-invasive diagnosis of fibrotic NASH with AUROC at 0.847 ⁇ 0.030 in the validation set. Furthermore, MACK-3 showed a significantly higher accuracy than the combinations A, B and C previously described in WO2014/049131 , both in the derivation and in the validation sets. Table 15: AUROC of MACK-3 and combinations A, B and C for the diagnosis of fibrotic NASH
  • Example 1 The study was conducted on the population described in Example 1 , obtained by pooling the data from three published studies performed in Angers (Boursier J et al., J Hepatol. 2016 Sep;65(3):570-8), Antwerp (Francque SM et al., Clin Gastroenterol Hepatol 2012;10:1162-8) and Nice (Anty R et al., Aliment Pharmacol Ther 2010;32: 1315-22). Their characteristics are presented hereinabove in Example 1 and detailed in Table 1.
  • the 846 patients were 2: 1 randomly divided into a derivation set (564 patients) and a validation set (282 patients), with patient characteristics not being significantly different between the two sets, as shown hereinabove in Table 6. Blood parameters
  • MACK-3 is the combination of the three biomarkers HOMA, AST, and CK-18 in a binary logistic regression.
  • the CHA index is determined and used as described in Polyzos SA et al. Ann Hepatol. 2013 Sep-Oct;12(5):749-57, and corresponds to the following formula: CK-18 x HOMA x AST)/1000.
  • Table 16 below compares the performance of MACK-3 and the CHA index for the diagnosis of fibrotic NASH in both the derivation and the validation sets. As shown below, the overall diagnostic accuracy of MACK-3 is significantly higher than that of the CHA index both in the derivation set (93.6% of correctly classified patients vs 76.6%> of correctly classified patients respectively, p ⁇ 0.001) and in the validation set (93.3%> of correctly classified patients vs 75.3% of correctly classified patients, respectively, p ⁇ 0.001).
  • DA overall diagnostic accuracy ⁇ i.e., rate of well-classified patients, %); Se: sensitivity (%); Spe: specificity (%); NPV: negative predictive value (%); PPV: positive predictive value; -LR: negative likelihood ratio; +LR: positive likelihood ratio; OR: diagnostic odds ratio

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biotechnology (AREA)
  • Analytical Chemistry (AREA)
  • Cell Biology (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Communicable Diseases (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un procédé d'évaluation de la présence et/ou de la gravité d'une stéatohépatite non alcoolique fibreuse (NASH fibreuse) chez un sujet, comprenant la mesure de la cytokératine 18 (CK-18), de l'aspartate aminotransférase (AST) et l'un d'une évaluation de modèle d'homéostasie de la résistance à l'insuline (HOMA), d'un rapport glucose/insuline à jeun (FGIR) ou d'un indice quantitatif de sensibilité à l'insuline (QUICKI) dans un échantillon dudit sujet. La présente invention concerne un test sanguin précis non invasif pour diagnostiquer une NASH fibreuse comprenant la détermination d'un score basé sur la combinaison de CK-18, AST et l'un de HOMA, FGIR ou QUICKI. La présente invention concerne également un procédé de surveillance de la progression d'une NASH fibreuse et un procédé de surveillance de la réponse au traitement chez un sujet traité pour une NASH fibreuse.
PCT/EP2018/054809 2017-02-27 2018-02-27 Diagnostic non invasif de stéatohépatite non alcoolique fibreuse WO2018154140A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
ES18705960T ES2942418T3 (es) 2017-02-27 2018-02-27 Diagnóstico no invasivo de la esteatohepatitis no alcohólica fibrosante (NASH)
US16/486,884 US20200011869A1 (en) 2017-02-27 2018-02-27 Non-invasive diagnosis of fibrotic non-alcoholic steatohepatitis
EP18705960.5A EP3586141B8 (fr) 2017-02-27 2018-02-27 Diagnostic non-invasif de la stéatohépatite non alcoolique (nash) fibrosante

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201762464106P 2017-02-27 2017-02-27
US62/464,106 2017-02-27
EP17305615 2017-05-24
EP17305615.1 2017-05-24

Publications (1)

Publication Number Publication Date
WO2018154140A1 true WO2018154140A1 (fr) 2018-08-30

Family

ID=59054049

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2018/054809 WO2018154140A1 (fr) 2017-02-27 2018-02-27 Diagnostic non invasif de stéatohépatite non alcoolique fibreuse

Country Status (1)

Country Link
WO (1) WO2018154140A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020208549A1 (fr) * 2019-04-12 2020-10-15 Metadeq Limited Nouveau marqueur pathologique et ses utilisations

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2490026A1 (fr) 2009-10-16 2012-08-22 Mochida Pharmaceutical Co., Ltd. Marqueur associé à la stéatohépatite non alcoolique
US20130183687A1 (en) 2007-06-14 2013-07-18 Zobair M. YOUNOSSI Methods of diagnosing non-alcoholic steatohepatitis (nash)
WO2014049131A1 (fr) 2012-09-28 2014-04-03 Université d'Angers Test sanguin précis pour le diagnostic non invasif de la stéatohépatite non alcoolique
WO2015192854A2 (fr) 2014-06-16 2015-12-23 Aarhus Universitet Méthodes et outils de prédiction d'une stéatose hépatique

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130183687A1 (en) 2007-06-14 2013-07-18 Zobair M. YOUNOSSI Methods of diagnosing non-alcoholic steatohepatitis (nash)
EP2490026A1 (fr) 2009-10-16 2012-08-22 Mochida Pharmaceutical Co., Ltd. Marqueur associé à la stéatohépatite non alcoolique
WO2014049131A1 (fr) 2012-09-28 2014-04-03 Université d'Angers Test sanguin précis pour le diagnostic non invasif de la stéatohépatite non alcoolique
WO2015192854A2 (fr) 2014-06-16 2015-12-23 Aarhus Universitet Méthodes et outils de prédiction d'une stéatose hépatique

Non-Patent Citations (28)

* Cited by examiner, † Cited by third party
Title
ALBERTI KG ET AL., CIRCULATION, vol. 120, 2009, pages 1640 - 1645
ANGULO P ET AL., HEPATOLOGY, vol. 45, 2007, pages 846 - 54
ANTY R ET AL., ALIMENT PHARMACOL THER, vol. 32, 2010, pages 1315 - 22
BOURSIER ET AL., J HEPATOL., vol. 65, no. 3, September 2016 (2016-09-01), pages 570 - 8
BOURSIER J ET AL., J HEPATOL, vol. 65, no. 3, September 2016 (2016-09-01), pages 570 - 8
BOURSIER J ET AL., J HEPATOL., vol. 65, no. 3, September 2016 (2016-09-01), pages 570 - 8
BRUNT EM ET AL., HEPATOLOGY, vol. 53, 2011, pages 810 - 20
CALES P ET AL., CLIN BIOCHEM, vol. 41, 2008, pages 10 - 18
CHALASANI ET AL., HEPATOLOGY, vol. 55, no. 6, 2012, pages 2005 - 2023
CHALASANI N ET AL., GASTROENTEROLOGY, vol. 142, 2012, pages 1592 - 609
FLORIANE RUDWILL ET AL: "Effect of enforced physical inactivity induced by 60-day of bed rest on hepatic markers of NAFLD in healthy normal-weight women", LIVER INTERNATIONAL, vol. 35, no. 6, 21 June 2015 (2015-06-21), GB, pages 1700 - 1706, XP055391307, ISSN: 1478-3223, DOI: 10.1111/liv.12743 *
FORTHOFER RN; LEE ES; HERNANDEZ M: "Biostatistics: A guide to Design, Analysis, and Discovery", 2006, ELSEVIER
FRANCQUE SM ET AL., CLIN GASTROENTEROL HEPATOL, vol. 10, 2012, pages 1162 - 8
HAMZA RT1 ET AL: "Serum Pentraxin 3 Fragment as a Noninvasive Marker of Nonalcoholic Fatty Liver Disease in Obese Children and Adolescents", HORMONE RESEARCH IN PAEDIATRICS, S. KARGER AG, CH, vol. 86, no. 1, 17 June 2016 (2016-06-17), pages 11 - 20, XP009504148, ISSN: 1663-2818, DOI: 10.1159/000446566 *
HARRISON SA ET AL., GUT, vol. 57, 2008, pages 1441 - 7
J HEPATOL, vol. 64, 2016, pages 1388 - 1402
KATZ A ET AL., J CLIN ENDOCRINOL METAB, vol. 85, 2000, pages 2402 - 2410
KLEINER DE ET AL., HEPATOLOGY, vol. 41, 2005, pages 1313 - 21
MATTHEWS ET AL., DIABETOLOGIA, vol. 28, 1985, pages 412 - 419
POLYZOS SA ET AL., ANN HEPATOL, vol. 12, no. 5, September 2013 (2013-09-01), pages 749 - 57
POLYZOS SA ET AL., ANN HEPATOL., vol. 12, no. 5, September 2013 (2013-09-01), pages 749 - 57
SANYAL AJ ET AL., HEPATOLOGY, vol. 54, 2011, pages 344 - 53
SANYAL AJ ET AL., HEPATOLOGY, vol. 61, 2015, pages 1392 - 405
SHAH AG ET AL., CLIN GASTROENTEROL HEPATOL, vol. 7, 2009, pages 1104 - 12
SHROUT PE; FLEISS JL, PSYCHOL BULL, vol. 86, 1979, pages 420 - 428
STERGIOS A POLYZOS ET AL: "Adipocytokines and cytokeratin-18 in patients with nonalcoholic fatty liver disease: Introduction of CHA index Correspondence and reprint request", 1 September 2013 (2013-09-01), pages 749 - 757, XP055391300, Retrieved from the Internet <URL:http://www.annalsofhepatology.com/revista/numeros/2013/06_135_v12n5_2013_AdipocytokinesCytokeratin.pdf> [retrieved on 20170717] *
STERLING RK ET AL., HEPATOLOGY, vol. 43, 2006, pages 1317 - 25
YOUNOSSI ZM ET AL., HEPATOLOGY, vol. 64, 2016, pages 73 - 84

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020208549A1 (fr) * 2019-04-12 2020-10-15 Metadeq Limited Nouveau marqueur pathologique et ses utilisations
CN113785202A (zh) * 2019-04-12 2021-12-10 美泰达有限公司 新病理标志物及其用途

Similar Documents

Publication Publication Date Title
Mantovani et al. Association between non-alcoholic fatty liver disease and bone turnover biomarkers in post-menopausal women with type 2 diabetes
Vos et al. NASPGHAN clinical practice guideline for the diagnosis and treatment of nonalcoholic fatty liver disease in children: recommendations from the Expert Committee on NAFLD (ECON) and the North American Society of Pediatric Gastroenterology, Hepatology and Nutrition (NASPGHAN)
Nickolas et al. Discriminants of prevalent fractures in chronic kidney disease
US10861582B2 (en) Non-invasive method for assessing liver fibrosis progression
Lacquaniti et al. Apelin and copeptin: two opposite biomarkers associated with kidney function decline and cyst growth in autosomal dominant polycystic kidney disease
US20180098728A1 (en) Non-invasive method for assessing the presence or severity of liver fibrosis based on a new detailed classification
Hwang et al. Urinary periostin excretion predicts renal outcome in IgA nephropathy
JP6062898B2 (ja) ガン罹患の予測のためのバイオマーカー
Moe et al. Association of hepatitis C virus infection with prevalence and development of kidney disease
Chan et al. Aldosterone level after saline infusion test could predict clinical outcome in primary aldosteronism after adrenalectomy
Preyer et al. Serum uric acid is associated with incident hip fractures in women and men–Results from a large Austrian population-based cohort study
Tavaglione et al. Head‐To‐Head Comparison Between Phosphatidylethanol Versus Indirect Alcohol Biomarkers for Diagnosis of MetALD Versus MASLD: A Prospective Study
Alkurt et al. The relationship between papillary thyroid cancer and triglyceride/glucose index, which is an indicator of insulin resistance.
Kumar et al. A clinical study of insulin resistance in patients with chronic pancreatitis
EP3586141B1 (fr) Diagnostic non-invasif de la stéatohépatite non alcoolique (nash) fibrosante
Kwon et al. Cystatin C is better than serum creatinine for estimating glomerular filtration rate to detect osteopenia in chronic kidney disease patients
Zayed et al. Serum levels of carcinoembryonic antigen in patients with type 2 diabetes
WO2018154140A1 (fr) Diagnostic non invasif de stéatohépatite non alcoolique fibreuse
Erkan et al. The relationship between insulin resistance, metabolic syndrome and nonalcoholic fatty liver disease in non-obese non-diabetic Turkish individuals: A pilot study
Choi et al. A composite Blood biomarker including AKR1B10 and cytokeratin 18 for progressive types of nonalcoholic fatty liver disease
Yang et al. Fasting plasma glucose levels predict steroid-induced abnormal glucose metabolism in patients with non-diabetic chronic kidney disease: a prospective cohort study
Home Noninvasive tests (NITs) in the management of nonalcoholic steatohepatitis (NASH)
Greco et al. Kinetics of the cell cycle arrest biomarkers (TIMP2 and IGFBP7) for the diagnosis of acute kidney injury in critically ill COVID-19 patients. Diagnostics (Basel). 2023; 13: 317
Ratziu et al. Aramchol improves hepatic fibrosis in MASH: Results of multimodality assessment using both conventional and digital pathology
Iqbal et al. The Predictive Potential of C-Peptide in Differentiating Type 1 Diabetes From Type 2 Diabetes in an Outpatient Population in Abu Dhabi

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18705960

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2018705960

Country of ref document: EP

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载