Model and device for predicting HBsAg clearance after hematopoietic stem cell transplantation in HBsAg-positive patients

Abstract

This invention discloses a predictive model and device for predicting HBsAg clearance after hematopoietic stem cell transplantation in HBsAg-positive patients. This invention retrospectively collected HBV test results from HBsAg-positive patients who underwent allo-HSCT at Peking University People's Hospital from January 2010 to December 2020, and established a predictive model CSE for HBsAg clearance outcomes in HBsAg-positive patients after allo-HSCT. Using this model, based on data regarding the presence or absence of CMV in the HBsAg-positive patient after hematopoietic stem cell transplantation, and whether the hematopoietic stem cell donor is HBsAg-positive and HBeAb-positive, the HBsAg clearance outcome in HBsAg-positive patients after allo-HSCT can be effectively predicted and assessed clinically.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a model and device for predicting HBsAg clearance after hematopoietic stem cell transplantation in HBsAg-positive patients. Background Technology

[0002] Hepatitis B virus (HBV) is the pathogen that causes hepatitis B (HBV for short), belonging to the family Hepatoviridae. HBV infection is a global public health problem. Because the clearance of hepatitis B surface antigen (HBsAg) is closely related to the occurrence of complications and prognosis of HBV infection, it is generally considered the ideal treatment target for patients with chronic HBV infection. However, HBsAg clearance is not common with current treatments (antiviral drugs and interferon).

[0003] Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is one of the most effective treatments for various hematological diseases. For patients with chronic HBV infection, the prognosis after allo-HSCT has been extensively studied and reported due to liver-related complications. However, the impact of allo-HSCT on the efficacy of treatment for chronic HBV infection remains unclear. It has been reported that some chronic HBV patients have experienced HBsAg loss after allo-HSCT, possibly due to the shift in humoral and cellular immunity. Therefore, allo-HSCT has the potential to help cure chronic HBV infection and even help clear HBsAg. However, due to the limited number of relevant studies, this hypothesis still requires further verification. Summary of the Invention

[0004] The technical problem to be solved by this invention is how to predict the HBsAg clearance outcome after hematopoietic stem cell transplantation in HBsAg-positive patients.

[0005] To address the aforementioned technical problems, this invention first provides an apparatus for predicting HBsAg clearance outcomes after hematopoietic stem cell transplantation in HBsAg-positive patients. The apparatus includes a data receiving module and a data processing module. The data receiving module is configured to receive data on whether the HBsAg-positive patient undergoing hematopoietic stem cell transplantation has CMV (Chronic Virus Vulnerability), whether the donor's HBsAb (HbsAb) is positive, and whether the donor's HBeAb (HBeAb) is positive. The data processing module processes the data using a model called CSE (Contingent Sequence of Predictive Study) to predict the HBsAg clearance risk after hematopoietic stem cell transplantation in HBsAg-positive patients, outputting the HBsAg clearance outcome for the HBsAg-positive patient after hematopoietic stem cell transplantation.

[0006] The hematopoietic stem cell transplantation described in this article can be allogeneic hematopoietic stem cell transplantation (allo-HSCT).

[0007] In the above-described apparatus, the data processing module can be established through a method including the following steps:

[0008] A1-1) Obtain the risk score for the HBsAg-positive patient to be tested: Record 1 point for HBsAg-positive patients without CMV blood after transplantation, otherwise (with CMV blood) is recorded as 0 points; 1 point for HBsAg-positive patients whose allogeneic hematopoietic stem cell donor HBbsAb is positive (represented by donor HBsAb(+) in Table 1), otherwise (donor HBsAb is negative) is recorded as 0 points; 1 point for HBsAg-positive patients whose allogeneic hematopoietic stem cell donor HBeAb is positive (represented by donor HBeAb(+) in Table 1), otherwise (donor HBeAb is negative) is recorded as 0 points.

[0009] A1-2) Calculate the risk value of HBsAg clearance in HBsAg-positive patients to be tested: sum the risk scores to obtain the total score, and the total score is the risk value;

[0010] A1-3) Output the HBsAg risk grouping results: if the risk value is greater than or equal to 0 and less than or equal to 1, the risk grouping result is HBsAg clearing low-risk group; if the risk value is equal to 2, the risk grouping result is HBsAg clearing medium-risk group; if the risk value is equal to 3, the risk grouping result is HBsAg clearing high-risk group.

[0011] A1-4) Output the HBsAg clearance results of the HBsAg positive patients to be tested: The HBsAg clearance rate of the HBsAg positive patients to be tested in the high-risk group is higher than that in the low-risk group and the medium-risk group.

[0012] To address the aforementioned technical problems, this invention also provides the application of substances for detecting CMV serocongestion in the preparation of products that predict HBsAg clearance outcomes after hematopoietic stem cell transplantation in HBsAg-positive patients.

[0013] To address the aforementioned technical problems, this invention also provides the application of substances for detecting HBsAgAb and HbeAb in the preparation of products that predict HBsAg clearance outcomes after hematopoietic stem cell transplantation in HBsAg-positive patients.

[0014] The HbsAb and HbeAb can be the HbsAb and HbeAb of the hematopoietic stem cell transplant donor.

[0015] To address the aforementioned technical problems, this invention also provides the application of substances for detecting CMV serocongestion, substances for detecting HBsAgAb, and substances for detecting HbeAb in the preparation of products that predict HBsAg clearance outcomes after hematopoietic stem cell transplantation in HBsAg-positive patients.

[0016] In the above applications, the substances for detecting HBsAb and HBeAb can be instruments for detecting serological indicators. The substance for detecting CMV seroprevalence can be a CMV PCR kit.

[0017] The HBsAb and HBeAb can be the HBsAb and HBeAb from a hematopoietic stem cell transplant donor. The CMV PCR kit can be Abbott CMV PCR.

[0018] The instrument used to detect serological indicators can be the Abbott AxSYM fully automated immunoassay analyzer.

[0019] In the above applications, the product may be a reagent, a kit, and / or a device.

[0020] To address the aforementioned technical problems, this invention also provides a kit for predicting HBsAg clearance outcomes after hematopoietic stem cell transplantation in HBsAg-positive patients. The kit includes a parameter detection device; this device is used to detect whether the HBsAg-positive patient undergoing hematopoietic stem cell transplantation has CMV (Chronic Virus Vulnerability) and to detect whether the donor of the hematopoietic stem cells is HBsAg-positive and whether the donor's HBeAb (Hyperthyroidism-related antigen) is HBsAg-positive.

[0021] The kit described above may also include a readable carrier. The readable carrier may contain the following information:

[0022] HBsAg-positive patients without CMV serology after transplantation are scored 1 point; otherwise (with CMV serology), they are scored 0 points.

[0023] For HBsAg-positive patients undergoing transplantation, a score of 1 is awarded if the donor's HBsAb is positive (represented by donor HBsAb(+) in Table 1), and 0 is awarded if the donor's HBsAb is negative.

[0024] For HBsAg-positive patients undergoing transplantation, a score of 1 is awarded if the donor's HBeAb is positive (represented by donor HBeAb(+) in Table 1), and 0 is awarded if the donor's HBeAb is negative.

[0025] The total score of HBsAg-positive patients after transplantation is recorded and output as a readable vector. When the output of the readable vector is 0-1, the HBsAg-positive patient is in the low-risk group for HBsAg clearance after hematopoietic stem cell transplantation; when the output of the readable vector is 2, the HBsAg-positive patient is in the intermediate-risk group for HBsAg clearance after hematopoietic stem cell transplantation; when the output of the readable vector is 3, the HBsAg-positive patient is in the high-risk group for HBsAg clearance after hematopoietic stem cell transplantation. The HBsAg clearance rate of the high-risk group patients after hematopoietic stem cell transplantation may be higher than that of the intermediate-risk group patients and the low-risk group patients.

[0026] To address the aforementioned technical problems, the present invention also provides a computer-readable storage medium storing a computer program that causes a computer to perform the following steps:

[0027] B1) Input data into the computer regarding whether the HBsAg-positive patient has CMV after hematopoietic stem cell transplantation, whether the donor of the hematopoietic stem cells is positive for HBsAg and whether the donor is positive for HBeAb.

[0028] B2) Convert the data into a risk value for HBsAg clearance after hematopoietic stem cell transplantation in the HBsAg-positive patient to be tested, and predict the HBsAg clearance outcome after hematopoietic stem cell transplantation in the HBsAg-positive patient to be tested based on the risk value.

[0029] In the aforementioned computer-readable storage medium, the risk value can be obtained by a method including the following steps:

[0030] B2-1) Obtain the risk score for the HBsAg-positive patient to be tested: Record 1 point for HBsAg-positive patients without CMV emia after transplantation, otherwise (with CMV emia) is recorded as 0 points; 1 point for HBsAg-positive patients whose allogeneic hematopoietic stem cell donor HBbsAb is positive (represented by donor HBsAb(+) in Table 1), otherwise (donor HBsAb is negative) is recorded as 0 points; 1 point for HBsAg-positive patients whose allogeneic hematopoietic stem cell donor HBeAb is positive (represented by donor HBeAb(+) in Table 1), otherwise (donor HBeAb is negative) is recorded as 0 points.

[0031] B2-2) Calculate the risk value of HBsAg clearance in HBsAg-positive patients: sum the risk scores to obtain the total score, and the total score is the risk value.

[0032] In the computer-readable storage medium described above, the computer program may also cause the computer to perform the following steps:

[0033] B3-1) Output the HBsAg risk grouping results: if the risk value is greater than or equal to 0 and less than or equal to 1, the risk grouping result is HBsAg clearing low-risk group; if the risk value is equal to 2, the risk grouping result is HBsAg clearing medium-risk group; if the risk value is equal to 3, the risk grouping result is HBsAg clearing high-risk group.

[0034] B3-2) Output the HBsAg clearance results of the HBsAg positive patients to be tested: The HBsAg clearance rate of the HBsAg positive patients to be tested in the high-risk group is higher than that in the low-risk group and the medium-risk group.

[0035] Given the relatively limited data on HBsAg clearance after allo-HSCT in patients with chronic HBV infection, this invention retrospectively collected HBV test results from HBsAg-positive patients who underwent allo-HSCT at Peking University People's Hospital from January 2010 to December 2020. The study investigated the incidence and risk factors of HBsAg clearance, and explored the therapeutic effect of HSCT on HBV infection. Furthermore, this invention established a predictive model for HBsAg clearance outcomes in HBsAg-positive patients after allo-HSCT to better assess the HBsAg clearance results after allo-HSCT. Attached Figure Description

[0036] Figure 1 Performance evaluation of the "CSE" prediction model. a) ROC curve for the development cohort; b) ROC curve for the validation cohort. The vertical axis represents sensitivity, and the horizontal axis represents 1-specificity.

[0037] Figure 2 Calibrate the performance of the "CSE" prediction model. a) is the calibration curve for the development cohort; B = 1000 repetitions, boot; Mean absolute error = 0.046, n = 83; b) is the calibration curve for the validation cohort. The vertical axis represents the probability of observation, and the horizontal axis represents the probability of prediction.

[0038] Figure 3 DCA analysis for the "CSE" prediction model. a) is the decision curve for the development cohort; b) is the decision curve for the validation cohort. The vertical axis represents the net benefit rate, and the horizontal axis represents the threshold probability. Detailed Implementation

[0039] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0040] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0041] The study type in this embodiment of the invention is a retrospective observational study.

[0042] The research objects in this embodiment of the invention are as follows:

[0043] Inclusion criteria: Patients who underwent allogeneic hematopoietic stem cell transplantation (hereinafter referred to as transplantation) at the Department of Hematology, Peking University People's Hospital between January 2010 and December 2020 and were HBsAg positive in pre-transplant screening.

[0044] Exclusion criteria: age <18 or >70 years; coexisting viral hepatitis; missing hepatitis B test data; patients with severe organ complications or other autoimmune diseases; or patients deemed unsuitable for the study by the investigators.

[0045] The grouping of the research objects in this embodiment of the invention is as follows:

[0046] A retrospective review of electronic medical records was conducted to include HBsAg-positive patients who underwent allo-HSCT between January 2010 and December 2020. The HBsAg clearance status after allo-HSCT in HBsAg-positive patients was assessed from the cohort data, and the patients were divided into an HBsAg clearance group and an HBsAg non-clearance group for data collection and analysis.

[0047] The research steps and the selection and confirmation of the main measurement indicators or outcome indicators in this embodiment of the invention are as follows:

[0048] Using electronic medical records and institutional databases, data on demographics, pre-transplant hepatitis B serology and HBV-DNA quantification in donors and recipients, pre-transplant liver function, antiviral treatment status, time to HBsAg seroconversion in recipients after transplantation, HBV-DNA quantification, and whether hepatitis B surface antibody (HBsAb) became seropositive, as well as their titers, were extracted. The primary endpoint of this study was HBsAg clearance. Secondary endpoints included HBsAb production and HBV viral load quantification at the time of HBsAg clearance. HBsAg clearance was defined as two consecutive negative HBsAg tests lasting at least one week. HBsAg non-clearance was defined as persistent HBsAg positivity or re-positivity within one week of seroconversion.

[0049] Statistical analysis method of this invention embodiment:

[0050] For continuous variables, use mean ± standard deviation. The distribution was indicated, and the Kolmogorov-Smirnov test was used to determine if it was normally distributed. This invention employs Logistic regression to analyze risk factors, constructing a predictive model based on independent risk factors and their beta values. Statistical analysis was performed using GraphPad Prism version 6 (GraphPad Software, San Diego, CA), SPSS version 23.0 for Windows (IBM, Armonk, NY), and R 3.5.1 software. A p-value < 0.05 was considered statistically significant.

[0051] Example 1: Establishment of a model for predicting HBsAg clearance after hematopoietic stem cell transplantation in HBsAg-positive patients

[0052] According to the inclusion and exclusion criteria, a total of 125 patients who were HBsAg positive before allo-HSCT were included in the study. 89 patients were male and the remaining 36 were female. The median age was 39 (range 6-63 years) and the median follow-up time was 861.0 (range 36.0-4057.0) days post-transplant.

[0053] Of these, 62 HBsAg-positive patients (49.6%) achieved HBsAg clearance after allo-HSCT, with clearance time ranging from 354 days post-transplant (17-1877 days).

[0054] The remaining 63 HBsAg-positive patients did not clear HBsAg after allo-HSCT and belonged to the HBsAg non-clearance group.

[0055] Patients who underwent allo-HSCT transplantation between 2016 and 2020 (83 patients) were selected as the development cohort, and patients who underwent allo-HSCT transplantation between 2010 and 2015 (42 patients) were selected as the validation cohort. Logistic univariate and multivariate analysis revealed that donor HBsAg positivity (hepatitis B surface antibody), donor HBeAb positivity (hepatitis B e antibody), and absence of post-transplant CMV seroprevalence (cytomegalovirus (CMV) seroprevalence) were independent risk factors for HBsAg clearance after allo-HSCT in HBsAg-positive patients. Based on the independent risk factors and their β coefficients analyzed in the development cohort, a predictive model for HBsAg clearance after hematopoietic stem cell transplantation in HBsAg-positive patients, "CSE" (Table 1), was constructed.

[0056] The detection methods for donor HBsAb (HBsAb and HBeAb) were as follows: Serological markers of hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBsAb), hepatitis B core antibody (HBcAb), hepatitis B e antigen (HBeAg), and hepatitis B e antibody (HBeAb) were detected using the Abbott AxSYM (Abbott Diagnostics, Abbott Park, IL) fully automated immunoassay analyzer. The detection method for CMV seroprevalence was as follows: Abbott CMV PCR was used. The CMV-DNA in the patient's plasma was detected as specifically positive.

[0057] The “CSE” prediction model scores a total of 3 points:

[0058] HBsAg-positive patients who undergo allo-HSCT transplantation without CMV seremia are scored 1 point; otherwise (with CMV seremia) they are scored 0 points.

[0059] For HBsAg-positive patients undergoing allo-HSCT transplantation, a positive HBsAb donor (represented by donor HBsAb(+) in Table 1) is scored as 1 point, otherwise (donor HBsAb is negative) is scored as 0 points.

[0060] For HBsAg-positive patients undergoing allo-HSCT transplantation, a positive HBeAb from the donor (represented by donor HBeAb(+) in Table 1) is scored as 1 point, otherwise (donor HBeAb is negative) is scored as 0 points.

[0061] Based on the “CSE” prediction model score, patients were divided into three groups: low, medium, and high risk. Patients with scores of 0-1 were classified as low-risk patients after allo-HSCT hematopoietic stem cell transplantation and HBsAg clearance, patients with scores of 2 were classified as medium-risk patients after allo-HSCT hematopoietic stem cell transplantation and HBsAg clearance, and patients with scores of 3 were classified as high-risk patients after allo-HSCT hematopoietic stem cell transplantation and HBsAg clearance.

[0062] Table 1. Predictive model "CSE" for HBsAg clearance after allo-HSCT transplantation in HBsAg-positive patients.

[0063]

[0064] The results showed that in the development cohort, the clearance rates of HBsAg in the low-risk group (31 patients), the intermediate-risk group (33 patients), and the high-risk group (19 patients) were 12.9%, 39.4%, and 89.5%, respectively.

[0065] Based on the predictive model, patients in the validation cohort were scored: 13 in the low-risk group, 21 in the intermediate-risk group, and 8 in the high-risk group. The actual HBsAg clearance rates for the three groups were 23.1%, 81%, and 100%, respectively. Therefore, the HBsAg clearance rate predicted by the CSE model for high-risk HBsAg-positive patients after allo-HSCT transplantation was higher than that for low-risk and intermediate-risk patients. Consequently, patients in the low-risk and intermediate-risk groups were more likely to achieve HBsAg clearance after allo-HSCT transplantation from high-risk HBsAg-positive patients predicted by the CSE model.

[0066] Receiver operating characteristic (ROC) reflects the balance between sensitivity and specificity. The area under the ROC curve is an important indicator of test accuracy; the larger the area under the ROC curve, the greater the diagnostic value of the test.

[0067] Sensitivity (true positive rate): The percentage of actual HBsAg clearance that is correctly predicted as HBsAg clearance according to experimental standards. The higher the sensitivity, the better; the ideal sensitivity is 100%.

[0068] Specificity (true negative rate): The percentage of actual non-HBcAg clearance that is correctly predicted as non-HBcAg clearance according to the test criteria. The higher the specificity, the better, with an ideal specificity of 100%.

[0069] ROC curve analysis showed that the "CSE" prediction model demonstrated good discriminative power in both the development and validation cohorts. Figure 1 In the graph, a represents the ROC curve for the development cohort, with an AUC value of 0.819; b represents the ROC curve for the validation cohort, with an AUC value of 0.860. (Calibration plot) Figure 2 (a and b) and DCA decision curve ( Figure 3 Results a) and b) respectively demonstrate that the prediction model has good calibration efficiency and can be applied clinically to benefit patients.

[0070] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.

Claims

1. A device for predicting HBsAg clearance outcomes after hematopoietic stem cell transplantation in HBsAg-positive patients, characterized in that: The device includes a data receiving module and a data processing module. The data receiving module is configured to receive data on whether the HBsAg-positive patient undergoing hematopoietic stem cell transplantation has CMV seroprevalence, whether the donor of the hematopoietic stem cells is HBsAb positive, and whether the donor is HBeAb positive. The data processing module is used to output the HBsAg clearance result of the HBsAg-positive patient after hematopoietic stem cell transplantation by using the CSE model to predict the HBsAg clearance risk after hematopoietic stem cell transplantation. The data processing module is established through a method including the following steps: A1-1) Obtain the risk score for the HBsAg-positive patient to be tested: Record 1 point for HBsAg-positive patients without CMV serology after transplantation, otherwise record 0 points; 1 point for HBsAg-positive patients whose allogeneic hematopoietic stem cell donor HBbsAb is positive, otherwise record 0 points; 1 point for HBsAg-positive patients whose allogeneic hematopoietic stem cell donor HbeAb is positive, otherwise record 0 points. A1-2) Calculate the risk value of HBsAg clearance in HBsAg-positive patients to be tested: sum the risk scores to obtain the total score, and the total score is the risk value; A1-3) Output the HBsAg risk grouping result: the risk value is greater than or equal to 0 and less than or equal to 1, and the risk grouping result is output as HBsAg low-risk group. The risk value is equal to 2, and the risk grouping result is output as the HBsAg clearing medium risk group; The risk value is equal to 3, and the output of the risk grouping removal result is HBsAg removal of high-risk group; A1-4) Output the HBsAg clearance results of the HBsAg positive patients to be tested: The HBsAg clearance rate of the HBsAg positive patients to be tested in the high-risk group is higher than that in the low-risk group and the medium-risk group.

2. A computer-readable storage medium storing a computer program that causes a computer to perform the following steps: B1) Input data into the computer regarding whether the HBsAg-positive patient has CMV after hematopoietic stem cell transplantation, whether the donor of the hematopoietic stem cells is positive for HBsAg and whether the donor is positive for HBeAb. B2) Convert the data into a risk value for HBsAg clearance after hematopoietic stem cell transplantation in the HBsAg-positive patient to be tested, and predict the HBsAg clearance outcome after hematopoietic stem cell transplantation in the HBsAg-positive patient to be tested based on the risk value. The risk value is obtained through a method including the following steps: B2-1) Obtain the risk score for the HBsAg-positive patient to be tested: Record 1 point for HBsAg-positive patients without CMV emia after transplantation, otherwise record 0 points; 1 point for HBsAg-positive patients whose allogeneic hematopoietic stem cell donor HBbsAb is positive, otherwise record 0 points; 1 point for HBsAg-positive patients whose allogeneic hematopoietic stem cell donor HbeAb is positive, otherwise record 0 points. B2-2) Calculate the risk value of HBsAg clearance in HBsAg-positive patients: sum the risk scores to obtain the total score, and the total score is the risk value.

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