METHODS AND KITS FOR PREDICTING THE RISK OF HAVING OR DEVELOPPING HEPATOCELLULAR CARCINOMA IN PATIENTS SUFFERING FROM CIRRHOSIS

Abstract

Plasma levels of different sub-populations of microvesicles (endothelial, leukocyte, platelet and hepatocyte) were measured by flow cytometry or ELISA/filtration on blood samples from 125 patients with cirrhosis, for which 36 of them were diagnosed with HCC at inclusion. The inventors show that the levels of microvesicles of endothelial origin (CD62E+) could predict the occurrence of HCC in patients with cirrhosis. Therefore the present invention relates to a method for determining whether a patient suffering from cirrhosis is at risk of having or developing hepatocellular carcinoma comprising determining the level of endothelial-derived microvesicles (e.g. by flow cytometry) in a blood sample obtained from the patient.

Claims

1. A method for determining whether a patient suffering from cirrhosis is at risk of having or developing hepatocellular carcinoma, comprising determining the level of endothelial-derived microvesicles in a blood sample obtained from the patient, wherein when the level of endothelial-derived microvesicles is higher than a predetermined reference value, the patient is at risk of having or developing hepatocellular carcinoma and when the level of endothelial-derived microvesicles is lower than the predetermined reference value, the patient is not at risk of having or developing hepatocellular carcinoma.

2. The method of claim 1 further comprising collecting a population of microvesicles from the blood sample and then contacting said population of microvesicles at least one binding partner directed against the specific surface markers of endothelial-derived microvesicles.

3. The method of claim 2 wherein the at least one binding partner is a monoclonal antibody.

4. The method of claim 3 wherein the monoclonal antibody is specific for CD62E.

5. The method of claim 2 wherein the level of endothelial-derived microvesicles is determined by flow cytometry.

6. The method of claim 1 wherein the predetermined reference value is established in a population of patients who do not have and/or have not developed hepatocellular carcinoma.

7. (canceled)

8. A method for determining whether a patient suffering from cirrhosis is at risk of having or developing hepatocellular carcinoma and treating and/or monitoring a patient identified as at risk, comprising i) determining the level of endothelial-derived microvesicles in a blood sample obtained from the patient, wherein when the level of endothelial-derived microvesicles is higher than a predetermined reference value, the patient is at risk of having or developing hepatocellular carcinoma and when the level of endothelial-derived microvesicles is lower than the predetermined reference value, the patient is not at risk of having or developing hepatocellular carcinoma, and ii) treating and/or monitoring a patient whose measurement is indicative of being at risk of having or developing hepatocellular carcinoma.

9. The method of claim 1, wherein the step of treating includes resecting the liver or the patient or transplanting a liver into the patient.

10. The method of claim 1, wherein the step of monitoring includes conducting ultrasonography of the patient every 3 months.

Description

FIGURE

[0023] FIG. 1: Cumulative risk of developing hepatocellular carcinoma in patients with cirrhosis according to circulating levels of endothelial microvesicles (CD62E+).

EXAMPLE

[0024] Plasma levels of different sub-populations of microvesicles (endothelial, leukocyte, platelet and hepatocyte) were measured by flow cytometry or ELISA/filtration on blood samples from 125 patients with cirrhosis, including 36 with a diagnosed with HCC at inclusion. Among the 89 patients without HCC diagnosed at inclusion and monitored prospectively by scan and/or ultrasound for 12 months, 15 developed HCC during follow-up. The capacity of circulating levels of microvesicles to predict the occurrence of HCC has been tested by the actuarial Kaplan Meier method. Results are expressed as median (interquartile range) or number of patients (%).

[0025] The 36 patients with active HCC at inclusion had less severe cirrhosis than the 89 patients without HCC [MELD at 11 (7-25) vs. 13 (10-16); P=0.027]. In order to determine whether the existence of HCC affected the levels of plasma microvesicles independently of severity and cause of cirrhosis, we performed a case-control study. We have matched the cause of cirrhosis and the severity of cirrhosis 21 of the 36 patients with HCC at inclusion with 21 of the 89 patients not having HCC at inclusion. The only difference between the 2 groups was that the rate of platelet-derived microvesicles (CD41+) slightly higher in the 21 patients with cirrhosis with HCC [729 (416-1688) vs. 606 (279-1566); P=0.026]. We then analyzed the impact of the size of the HCC nodule on the levels of microvesicles in the 36 patients with HCC. We did not see any difference between patients with a tumor volume greater than 5 cm3 and those with a volume below this limit. We then analyzed the ability of microvesicle levels to predict the occurrence of HCC in 89 patients without HCC at inclusion. Patients who developed HCC in the follow-up had 2-fold higher rates of endothelial microvesicles (CD62E+, p=0.032) microvesicle than patient who did not develop the cancer. The actuarial risk to develop HCC was 26% at 12 months in patients having level of microvesicles of endothelial origin in the 3rd tertile vs. 11% in others (log rank, p=0.03).

[0026] Our results show that the levels of microvesicles are not very different in patients with and without HCC when the tumor is small, a situation where there is a clinical need. In contrast, microvesicles of endothelial origin (CD62E+) could predict the occurrence of HCC in patients with cirrhosis.

REFERENCES

[0027] Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.