Search

CN-122029292-A - Method for screening patients with at-risk metabolic dysfunction-related steatohepatitis (MASH)

CN122029292ACN 122029292 ACN122029292 ACN 122029292ACN-122029292-A

Abstract

Provided herein are methods of screening patients with at-risk MASH (metabolic dysfunction-associated steatohepatitis, previously known as non-alcoholic steatohepatitis or NASH). Combining the levels of five circulating markers (i.e., alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), platelets, hsa-miR-34a and YKL-40), and finally the age and sex of the subject, allows for accurate identification of MASH at risk for non-liver cirrhosis. The information obtained from the levels of these markers also allows for screening of subjects as recipients or non-recipients of and/or categorizing subjects as responders or non-responders to treatment with MASH, in compliance with liver biopsy conditions.

Inventors

  • Y. Haji
  • J. Magnan Ensi

Assignees

  • 基恩菲特公司

Dates

Publication Date
20260512
Application Date
20241009
Priority Date
20231010

Claims (15)

  1. 1. A method for diagnosing a subject with a MASH at risk of non-liver cirrhosis, and/or for screening a subject for compliance with liver biopsy conditions, and/or for classifying a subject as a recipient or non-recipient of MASH treatment, and/or for classifying a subject as a responder or non-responder to MASH treatment, wherein the method comprises: Providing the level of alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), platelets, hsa-miR-34a and YKL-40 in a biological fluid sample of said subject, -Comparing the levels of ALAT, ASAT and platelets with reference levels obtained in samples from subjects suffering from MASH at risk of liver cirrhosis and comparing the levels of hsa-miR-34a and YKL-40 with reference levels obtained in samples from subjects not suffering from MASH at risk.
  2. 2. The method according to claim 1, wherein the method is for screening a subject for compliance with liver biopsy conditions in a MASH clinical trial.
  3. 3. The method of claim 1 or 2, further comprising determining the age and sex of the subject.
  4. 4. A method according to claim 3, Wherein the provided levels of ALAT, ASAT, platelet and age are combined in a first mathematical function to calculate a first score (S1), and Wherein the levels of hsa-miR-34a and YKL-40 provided are combined with the gender in a second mathematical function to calculate a second score (S2).
  5. 5. The method according to claim 4, wherein the first score is compared with a first cut-off value (co 1) and the second score is compared with a second cut-off value (co 2), Wherein said co1 is calculated from reference levels of ALAT, ASAT and platelets obtained from a sample from a MASH subject at risk of liver cirrhosis and the age of said subject, and said co2 is calculated from reference levels of hsa-miR-34a and YKL-40 obtained from a sample from a subject not suffering from MASH at risk and the sex of said subject, Wherein a subject having a first score lower than said co1 and a second score higher than said co2 is identified as a subject having a MASH at risk of non-liver cirrhosis, and/or a subject meeting liver biopsy conditions, and/or a subject being a recipient of MASH treatment, and/or a subject being a responder to MASH treatment.
  6. 6. The method according to claim 4 or 5, wherein the first score (S1) is obtained as a mathematical function defined as age (years) x ASAT (U/L)/platelets (x 10 9 /L)/ALAT (U/L).
  7. 7. The method according to claim 6, wherein the first cut-off value is comprised between 2 and 3, in particular comprised between 2.4 and 2.8, more in particular equal to 2.48 or 2.67, still more in particular equal to 2.48.
  8. 8. The method according to claim 4 or 5, wherein the second score (S2) is obtained from a logic function: Wherein the method comprises the steps of y = β0 + β1 Log10 (miR-34 a-5p (fold)) +β2 log10(YKL-40 (ng/ml)) + β3 Gender + beta 4 Log10 (miR-34 a-5p (fold)) Sex (sex) Wherein the method comprises the steps of If the subject is female, the sex is 0, or if the subject is male, the sex is 1, Beta 0 is comprised between-3 and 3, in particular between-2 and 2, Β1 is comprised between 1 and 5, in particular between 2 and 4, Beta.2 is comprised between 0 and 4.5, in particular between 0.5 and 3, Beta.3 is comprised between-2 and 2, in particular between-1 and 1, Beta.4 is comprised between-1 and 2, in particular between 0 and 2.
  9. 9. The method according to claim 8, wherein the second cut-off value is comprised between 0.4 and 0.8, in particular between 0.5 and 0.7, more in particular equal to 0.53 or 0.68, still more in particular equal to 0.53.
  10. 10. The method according to any one of claims 1 to 9, wherein the biological fluid sample is a blood sample or a blood derived sample, such as serum or plasma.
  11. 11. The method of any one of claims 1-10, wherein hsa-miR-34a is hsa-miR-34a-5p.
  12. 12. A kit comprising means for determining the levels of ALAT, ASAT, platelet, hsa-miR-34a-5p and YKL-40.
  13. 13. The kit of claim 12, comprising enzymatic reagents for measuring ALAT and ASAT, a cell counter for measuring platelets, an antibody directed against YKL-40, and primers and/or probes specific for hsa-miR-34a-5 p.
  14. 14. A computer program comprising instructions which, when executed by a processor/processing means, cause the processor/processing means to: a) Receiving the levels, age and sex of ALAT, ASAT, platelets, hsa-miR-34a, YKL-40 of the subject, B) (i) calculating a first score from said levels of ALAT, ASAT, platelets and said age, and (ii) calculating a second score from said levels of hsa-miR-34a and YKL-40 and said gender, C) (i) comparing the calculated first score with a first cutoff value, and (ii) comparing the calculated second score with a second cutoff value, D) If the first score is lower than the co1 and the second score is higher than the co2, assigning the subject to a MASH subject at risk of non-liver cirrhosis, and/or a patient meeting liver biopsy conditions, and/or a subject classified as a recipient of MASH treatment, and/or a group of subjects classified as a responder to MASH treatment, and if the first score is higher than the co1 and/or the second score is lower than the co2, the subject is not assigned to a group of MASH subjects at risk of non-liver cirrhosis, and/or a subject not meeting liver biopsy conditions, and/or a subject classified as a recipient of MASH treatment, and/or a subject classified as a responder to MASH treatment.
  15. 15. A computer-readable recording medium storing instructions for performing the method according to any one of claims 1 to 11.

Description

Method for screening patients with at-risk metabolic dysfunction-related steatohepatitis (MASH) The present invention relates to a method for screening patients suffering from MASH at risk. Background Metabolic dysfunction-associated steatohepatitis (MASH), previously known as nonalcoholic steatohepatitis or NASH, is a progressive condition that may lead to cirrhosis, hepatocellular carcinoma and end-stage liver disease. Has become the main indication of liver transplantation. Active steatohepatitis, which is histologically defined as metabolic dysfunction-related steatohepatitis (MASLD) activity score (MAS) 4 or more and significant fibrosis (F stage 2 or more), is commonly referred to as at-risk (at-risk) MASH, as it is associated with liver-related mortality, liver cancer and a higher risk of progression to cirrhosis (Harrison SA, et al 2020;5 (11): 970-985). The risk of liver-related mortality increases exponentially with increasing stages of fibrosis, and mortality and morbidity increase exponentially once cirrhosis progresses. In the absence of approved therapies, drug development to treat the at-risk MASH is an active area of research. Thus, patients with MASH at risk must be strictly selected for clinical trials with histological endpoints. However, patients with cirrhosis (F4) were excluded because the cirrhosis test had a specific design and endpoint (Noureddin M, et al, gastroenterology 2020; 159:422-427). Currently, patients who have included MASH at risk can only be examined by histopathology of Liver Biopsy (LB) specimens. However, liver biopsy has several recognized limitations, including sampling errors, inter-observer variability, and hospitalization. The main disadvantage is the significant risk of complications (including bleeding, pain and even death). Furthermore, the biopsy does not reflect changes in the whole liver, as it is a needle biopsy, and liver tissue obtained from the subintimal region does not represent the whole liver. Thus, biopsies do not constitute a reliable prognostic predictor. In addition, the use of LB in clinical trials presents significant logistical challenges such as surgical costs and significant screening failure rates (Sumida Y et al, world J Gastroenterol 2014; 20:475-485). Thus, screening methods are performed to select patients to be selected for liver biopsy. Stringent clinical trial inclusion and exclusion criteria can lead to narrow patient registration and limit the number of patients sent to subsequent liver biopsies while improving inclusion rate. Indeed, one of the biggest challenges of MASH assays is the very high rate of screening failure based on histological reasons, where LB does not confirm clinically suspected risky MASH (Ratziu V, et al, J hepatol 2022; 76:1263-1278). Generally, these results of LB cannot histologically confirm MASH at risk, and thus the screening failure rate increases. The high failure rate of screening not only means that many patients are unnecessarily exposed to the risks associated with LB, but also increases the number of patients to be screened and the overall cost of the trial. Furthermore, if the screening failure rate continues to be high, there is a risk of losing the aggressiveness of the investigation screening point for a particular trial. Performing a large number of liver biopsies also requires mobilization site specialists. Thus, in conducting a clinical trial of MASH, it is necessary to accurately identify patients with MASH at risk for non-cirrhosis and then conduct a Liver Biopsy (LB) for histological confirmation. To date, there is no non-invasive strategy for clinical trials of MASH to select patients for transfer of LB, other than relying on clinical and biological features that are neither sensitive nor specific. In fact, during clinical trial screening, a large number of patients were excluded after liver biopsy. This situation emphasizes the need to reduce the failure rate of Liver Biopsy (LBFR) to avoid the associated risks and costs. Therefore, there is a need to maximize the accuracy of screening methods to select subjects with MASH that are not at risk for cirrhosis to optimize the referral of LB. Furthermore, since patients with cirrhosis are not targeted by the developing MASH treatment at risk, it is also important and necessary to accurately diagnose patients with non-cirrhosis MASH at risk, which would be recipients and responders of these treatments in a real world environment outside the framework of clinical trials. Thus, there remains an unmet medical need to develop methods to identify patients with MASH at risk for non-cirrhosis, for referral to liver biopsies in MASH clinical trials, aimed at reducing the number of unnecessary procedures, reducing the failure rate of Liver Biopsies (LBFR), reducing the number of patients with cirrhosis that are referral to liver biopsies, reducing the cost of the screening procedure without increasing the number of screening required (NNS), and more