A Prospective, Single-arm Clinical Trial of Prevention of Severe Acute Graft-versus-host Disease After Adult Patients Receiving Allogeneic Hematopoietic Stem Cell Transplantation Using a daGOAT Model
Phase: PHASE2
Conditions: Transplant-Related Disorder
Sponsor: Institute of Hematology & Blood Diseases Hospital, China

 
Clinical study protocol 
Title: 
A prospective, single-arm clinical trial of prevention of severe acute 
graft-versus-host disease after adult patients receiving allogeneic 
hematopoietic stem cell transplantation using a daGOAT model 
Portocol 
number: 
daGOAT-adult-001 
Research 
unit： 
Institute of Hematology & Blood Diseases Hospital, Chinese 
Academy of Medical Sciences & Peking Union Medical College. 
Address： 
Nanjing Road, Heping district, Tianjin, China 
Version： 
5.0 
Version date： October 7, 2024 
 
 
 
 
Declaration of Secrecy 
 
This document is confidential information of Institute of Hematology & Blood Diseases Hospital, 
Chinese Academy of Medical Sciences & Peking Union Medical College (Tianjin, China) and is only 
used for the purpose of this clinical study. It shall not be disclosed to anyone other than the 
participating researchers and members of the institutional review board. This information cannot be 
used for any purpose other than the evaluation or implementation of clinical studies without the prior 
written consent of Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical 
Sciences & Peking Union Medical College. 
 
 

 
 
1 
1. 
Abstract 
 
Title 
A prospective, single-arm clinical trial of prevention of severe acute 
graft-versus-host disease after adult patients receiving allogeneic 
hematopoietic stem cell transplantation using the daGOAT model 
Study 
objective 
To evaluate the efficacy and safety of ruxolitinib for prophylactic 
therapy of adult patients who are predicted to have a high risk for 
developing severe acute graft-versus-host disease (aGVHD) by the 
daGOAT model. 
Study design This is a prospective single-arm historical-control clinical trial. Adult 
patients receiving human leukocyte antigen (HLA)-mismatched 
allogeneic hematopoietic stem cell transplantation (allo-HSCT) will 
be enrolled after October 1, 2022. The propensity score matching 
method will be used to select controls at a ratio of 1:3 from the 
historical cases. 
Study 
population 
Adult patients receiving HLA-mismatched allo-HSCT. 
Inclusion 
criteria 
1. Patients must be > 16 years of age; 
2. Patients receiving HLA-mismatched and non-cord blood allo-
HSCT; 
3. Patients who can take oral medication; 
4. Patients have to sign an informed consent form before the start of 
the research procedure. 
Exclusion 
criteria 
1. Tandem transplantation or multiple transplantations; 
2. Patients who are allergic to or cannot tolerate ruxolitinib; 
3. Mental or other medical conditions that make the patients unable 
to comply with the research treatment and monitoring 
requirements; 
4. Patients who are pregnant or cannot take appropriate contraceptive 
measures during treatment; 
5. Patients who are ineligible for the study due to other factors or will 
bear great risk if participating in the study. 
Sample size 
The incidence rate of severe aGVHD within 100 days of previous 
adult transplant patients at the Institute of Hematology & Blood 
Diseases Hospital, Chinese Academy of Medical Sciences & Peking 
Union Medical College (IHCAMS) was 18% during 1 December 

 
 
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2020–31 December 2021. The incidence rate after using the daGOAT 
model will fall to 8% per the researchers’ estimation. Comparison 
will be performed using a propensity score matching method to select 
controls at a ratio of 1:3 from the historical cases. To attain a 0.05 
significance level and a 0.8 power at a presumed 5% dropout rate, ≥ 
102 participants will need to be enrolled, and ≥ 306 historical 
controls will need to be matched. 
Dosage 
regimen 
 Model-predicted high-risk patients: ruxolitinib 5mg bid po until 
at least day 60 post-transplant and terminated after day 100. If 
severe hematological signs occur such as when there is severe 
neutropenia (<0.1×109/L), ruxolitinib can be used at half dose or 
discontinued as appropriate, and can continue to be used after 
hematology recovery. 
 Model-predicted moderate-risk patients: ruxolitinib 2.5mg bid p 
po until at least day 60 post-transplant and terminated after day 
100. If severe hematological signs occur such as when there is 
severe neutropenia (<0.1×109/L), ruxolitinib can be used at half 
dose or discontinued as appropriate, and can continue to be used 
after hematology recovery. 
 Model-predicted low risk: regular aGVHD prophylactic 
regimens. 
Study 
endpoints 
Primary endpoint: incidence of severe aGVHD after transplantation 
within 100 days. 
Secondary endpoints: incidence of aGVHD (any grade) in various 
target organs, overall survival after transplantation, relapse-free 
survival rate, relapse rate, incidence of severe infections, safety of 
treatment, and total cost of treatment. 
Safety 
evaluation 
Encompassing physical examination, vital signs, adverse events, 
concomitant therapy, and abnormal results of laboratory tests. 
Statistical 
analysis 
SAS and R statistical analysis software will be used to conduct the 
statistical analysis tailored to data properties. The propensity score 
matching method will be used to select controls at a ratio of 1:3 from 
the historical cases. Efficacy and safety variables will be compared 
between the two groups of the matched patients. All comparison tests 
will be two-sided, and statistical significance will be defined as P < 
0.05 in all the analyses. 

 
 
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2. 
Research background 
At present, there are about 40,000 new allogeneic hematopoietic stem cell 
transplantation (allo-HSCT) cases every year in the world [1, 2], among which 
about a quarter occur in China. Allo-HSCT has become the primary treatment 
method for some hematological diseases such as refractory and relapsed acute 
leukemia and bone marrow failure. The individuals are affected by dynamic and 
static multi-dimensional parameters as well as post-transplant immune 
reconstitution. The use of machine learning to integrate big data for predicting 
severe aGVHD (grade III–IV) may provide a new path towards understanding 
immune complications after transplantation. 
 
Previous foundation of research 
The researchers have compiled the postoperative clinical data of the patients who 
received HLA-mismatched allogeneic hematopoietic stem cell transplantation 
(allo-HSCT) at the IHCAMS from 2012 to 2021—named the ‘aGOAT dataset’, 
which contained 194 dynamic variables of 584 adult patients and 159 dynamic 
variables of 45 pediatric patients, including complete blood counts, blood 
biochemistry, electrolytes, cytokines, flow cytometry, antibody, fluid 
inflow/outflow, and vital signs. Severe aGVHD occurred within 100 days in 16% 
of the adult patients and 24% of the pediatric patients. A dynamic forecasting 
model for severe aGVHD, termed the ‘daGOAT model’, was constructed, 
achieving an AUROC score of more than 0.78. This result has been published in 
Nature Computational Science in 2022.[3] 
 
This study aims to prospectively evaluate the use of the daGOAT model in real-world 
clinical settings at the IHCAMS. 
 
1. 
Gratwohl, A., et al., One million haemopoietic stem-cell transplants: a retrospective observational study. 
Lancet Haematol, 2015. 2(3): p. e91-100. 

 
 
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2. 
Xu, L.P., et al., A review of hematopoietic cell transplantation in China: data and trends during 2008-
2016. Bone Marrow Transplant, 2017. 52(11): p. 1512-1518. 
3. 
Liu, X., et al. Dynamic forecasting of severe acute graft-versus-host disease after transplantation. Nat 
Comput Sci , 2022.2: p. 153-159. 
 
3. 
Study objective 
To evaluate the efficacy and safety of ruxolitinib for prophylactic therapy of adult 
patients who are predicted to have a high risk for developing severe aGVHD 
according to the daGOAT model. 
 
4. 
Study design 
This is a prospective single-arm historical-control clinical trial. Adult patients 
receiving HLA-mismatched allogeneic hematopoietic stem cell transplantation (allo-
HSCT) at the IHCAMS will be enrolled after October 1, 2022. The propensity score 
matching method will be used to select controls at a ratio of 1:3 from the historical 
cases. 
The daGOAT model will dynamically predict the risk for severe aGVHD daily from 
day 17 to day 23 after transplantation, and medication will be adjusted according to 
the predicted risk.  
The study flowchart is shown in Figure 1 (LR, low risk; MR, moderate risk; HR, high 
risk).  

 
 
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Figure 1. Study flowchart 
 
 
5. 
Study population 
Adult patients receiving HLA-mismatched allo-HSCT from the IHCAMS. 
 

 
 
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5.1 Inclusion criteria 
1) 
Patients must be > 16 years of age; 
2) 
Patients receiving HLA-mismatched and non-cord blood allo-HSCT; 
3) 
Patients who can take oral medication; 
4) 
Patients have to sign an informed consent form before the start of the 
research procedure. 
5.2 Exclusion criteria 
Patients who meet any of the following criteria will not be enrolled in the study: 
1) 
Tandem transplantation or multiple transplantations; 
2) 
Patients who are allergic to or cannot tolerate ruxolitinib; 
3) 
Mental or other medical conditions that make the patients unable to comply 
with the research treatment and monitoring requirements; 
4) 
Patients who are pregnant or cannot take appropriate contraceptive 
measures during treatment; 
5) 
Patients who are ineligible for the study due to other factors, or will bear 
great risk if participating in the study. 
5.3 Drop-out and withdrawal criteria 
1) 
Failure of neutrophil engraftment within 30 days of transplantation; 
2) 
A patient may withdraw from the study if he/she does not wish to continue 
participating in the study, and the date and reason for withdrawal shall be 
recorded. The investigator may also decide to discontinue a patient from 
the clinical study if there is an unacceptable risk. 
 
6. 
Dosage regimen 
1) Model-predicted high-risk patients: ruxolitinib 5mg bid po until at least day 
60 post-transplant and terminated after day 100. If severe hematological 
signs occur such as when there is severe neutropenia (<0.1×109/L), 
ruxolitinib can be used at half dose or discontinued until recovery. 

 
 
7 
2) Model-predicted moderate-risk patients: ruxolitinib 2.5mg bid. po until at 
least day 60 post-transplant and terminated after day 100. If severe 
hematological signs occur such as when there is severe neutropenia 
(<0.1×109/L), ruxolitinib can be used at half dose or discontinued until 
recovery  
3) 
Model-predicted low risk: regular aGVHD prophylactic regiments. 
 
7. 
Study assessment and follow-up 
7.1. Medical history 
Medical history (previous treatments, including chemotherapy). 
7.2. Physical examination 
Physical examination before transplantation, including body height, body 
weight, body surface area, and the Eastern Cooperative Oncology Group score. 
7.3. Laboratory tests  
Laboratory tests performed at diagnosis and before and after transplantation at 
various time points. 
1) 
Blood routine: Red blood cell count, hemoglobin, platelet count, white 
blood cell count, white blood cell classification count, etc. 
2) 
Urine routine: specific gravity, pH, urine sugar, urine protein, 
urobilinogen, ketone bodies, microscopic red blood cells, urine white 
blood cells, bacteria, crystals, tube type, etc. 
3) 
Coagulation function tests: activated partial thromboplastin time, plasma 
prothrombin time, international normalized ratio of prothrombin time, 
thrombin time, plasma fibrinogen, and plasma D-dimer. 
4) 
Bone marrow:  
 
Smear or biopsy analysis; 
 
Immunophenotype analysis by flow cytometry; 
 
Chromosome karyotype; 
 
Gene mutation analysis. 

 
 
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5) 
Blood biochemistry. 
 
Electrolyte includes sodium, potassium, chlorine, calcium and 
phosphorus, etc. 
 
Liver function tests include alanine aminotransferase, aspartate 
aminotransferase, alkaline phosphatase, γ-glutamyltransferase, lactate 
dehydrogenase, total protein, albumin, total bilirubin, direct bilirubin, 
total cholesterol, triglyceride, etc. 
 
Renal function tests include blood urea nitrogen, uric acid and 
creatinine, etc. 
6) 
Immune cell subset analysis by flow cytometry. 
7.4. Other examinations. 
1) 
12-lead electrocardiogram; 
2) 
Chest computed tomography; 
3) 
Abdominal ultrasound and abdominal CT when necessary; 
4) 
Echocardiography when necessary. 
5) 
Other examinations that the investigators consider appropriate. 
7.5. Follow-up. 
Patients will be followed up at days 14, 28, 42, 60, 90, 180, 270, 360 and 540 
after transplantation; data on infection, relapse, survival, and quality of life will 
be collected. 
 
8. 
Study endpoints 
8.1  Primary endpoint 
Incidence of severe aGVHD after transplantation within 100 days. 
8.2  Secondary endpoints 
Incidence of aGVHD (any grade) in various target organs, overall survival 
after transplantation, relapse-free survival rate, relapse rate, incidence of 
severe infections, safety of treatment, and total cost of treatment. 

 
 
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9. 
Adverse event and severe adverse event 
Starting from the date of signing the informed consent form until the last visit, all 
adverse events will be evaluated and recorded according to the Common Terminology 
Criteria for Adverse Events version 5.0. 
 
10. Sample size calculation 
The incidence rate of severe aGVHD within 100 days of previous adult transplant 
patients at the IHCAMS was 18% during 1 December 2020 ¬ 31 December 2021. The 
incidence rate after using the daGOAT model will fall to 8% per the researchers’ 
estimation. Comparison will be performed using a propensity score matching method 
to select controls at a ratio of 1:3 from the historical cases. To attain a 0.05 
significance level and a 0.8 power at a presumed 5% dropout rate, ≥ 102 
participants will need to be enrolled, and ≥ 306 historical controls will need to be 
matched. 
 
11. Statistical analysis 
SAS and R statistical analysis software will be used to conduct the statistical analysis 
tailored to data properties. The propensity score matching method will be used to 
select controls at a ratio of 1:3 from the historical cases. Efficacy and safety variables 
will be compared between the two groups of the matched patients. All comparison 
tests will be two-sided, and statistical significance will be defined as P < 0.05 in all the 
analyses. 
 
12. Ethical review 
The study will be conducted in accordance with the Declaration of Helsinki (2013), 
relevant regulations issued by the government of the People’s Republic of China, and 
additional precautions required by the ethics review committee at the IHCAMS. 

 
 
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Before the study, the investigator will obtain approvals from the ethics review 
committee at the IHCAMS regarding the study protocol data sheet, informed consent 
form, subject enrollment form, and other relevant information to be provided to the 
subject before enrollment. During the study, if there is any amendment to the study 
protocol data sheet, informed consent form, subject enrollment form, and other 
relevant information to be provided to the subject before enrollment, renewed 
approvals shall be obtained from the IHCAMS ethics review committee before 
continuation of the study. 
 
13. Preservation of research data 
All data of this study will be stored at the IHCAMS. Data sharing among the 
researchers will abide by the regulations of the People’s Republic of China regarding 
desensitization.