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PURPOSEPatients with relapsed or refractory T-cell acute lymphoblastic leukemia (r/r T-ALL) have few options and poor prognosis. The aim was to assess donor-derived anti-CD7 chimeric antigen receptor (CAR) T-cell safety and efficacy in patients with r/r T-ALL.METHODSIn this single-center, phase I trial, we administered anti-CD7 CAR T cells, manufactured from either previous stem-cell transplantation donors or new donors, to patients with r/r T-ALL, in single infusions at doses of 5 × 105or 1 × 106(±30%) cells per kilogram of body weight. The primary end point was safety with efficacy secondary.RESULTSTwenty participants received infusions. Adverse events including cytokine release syndrome grade 1-2 occurred in 90% (n = 18) and grade 3-4 in 10% (n = 2), cytopenia grade 3-4 in 100% (n = 20), neurotoxicity grade 1-2 in 15% (n = 3), graft-versus-host disease grade 1-2 in 60% (n = 12), and viral activation grade 1-2 in 20% (n = 4). All adverse events were reversible, except in one patient who died through pulmonary hemorrhage related to fungal pneumonia, which occurred at 5.5 months, postinfusion. Ninety percent (n = 18) achieved complete remission with seven patients proceeding to stem-cell transplantation. At a median follow-up of 6.3 months (range, 4.0-9.2), 15 remained in remission. CAR T cells were still detectable in five of five patients assessed in month 6, postinfusion. Although patients' CD7-positive normal T cells were depleted, CD7-negative T cells expanded and likely alleviated treatment-related T-cell immunodeficiency.CONCLUSIONAmong 20 patients with r/r T-ALL enrolled in this trial, donor-derived CD7 CAR T cells exhibited efficient expansion and achieved a high complete remission rate with manageable safety profile. A multicenter, phase II trial of donor-derived CD7 CAR T cells is in progress ( NCT04689659 ).

Pan and colleagues report one of the first prospective evaluations of planned sequential chimeric antigen receptor (CAR) T-cell therapy targeting CD19 and then CD22 in a phase 1 trial, indicating acceptable toxicity and encouraging durable efficacy in pediatric patients with relapsed/refractory (r/r) acute lymphoblastic leukemia (ALL).

Rationale: Viruses are the most common pathogens that can cause infection-related non-recurrent death after transplantation, occurring mostly from the early stages of hematopoietic stem cell transplantation (HSCT) to within 1 year after transplantation. Human coronavirus (HCoV)-NL63 is a coronavirus that could cause mortality among patients with underlying disease complications. Serological tests are of limited diagnostic value in immunocompromised hosts and cases of latent infection reactivation. In contrast, macro-genomic high-throughput (DNA and RNA) sequencing allows for rapid and accurate diagnosis of infecting pathogens for targeted treatment. Patient concerns: In this report, we describe a patient who exhibited acute B-lymphocytic leukemia and developed complicated pulmonary HCoV-NL63 infection after a second allogeneic HSCT (allo-HSCT). Six months after the second allo-HSCT, he developed sudden-onset hyperthermia and cough with decreased oxygen saturation. Chest computed tomography (CT) suggested bilateral multiple rounded ground-glass opacities with the pulmonary lobules as units. Diagnoses: HCoV-NL63 was detected by metagenomic next-generation sequencing (NGS), and HCoV-NL63 viral pneumonia was diagnosed. Interventions: The treatment was mainly based on the use of antiviral therapy, hormone administration, and gamma-globulin. Outcomes: After the therapy, the body temperature returned to normal, the chest CT findings had improved on review, and the viral copy number eventually became negative. Lessons: The latest NGS is an effective method for early infection diagnosis. The HCoV-NL63 virus can cause inflammatory factor storm and alter the neutrophil-to-lymphocyte ratio (NLR). This case suggests that the patient's NLR and cytokine levels could be monitored during the clinical treatment to assess the disease and its treatment outcome in a timely manner.

Despite worldwide promising clinical outcome of CD19 CAR-T therapy, relapse after this therapy is associated with poor prognosis and has become an urgent problem to be solved. We conducted a CD22 CAR T-cell therapy in 34 relapsed or refractory (r/r) B-ALL pediatric and adult patients who failed from previous CD19 CAR T-cell therapy. Complete remission (CR) or CR with incomplete count recovery (CRi) was achieved in 24 of 30 patients (80%) that could be evaluated on day 30 after infusion, which accounted for 70.5% of all 34 enrolled patients. Most patients only experienced mild cytokine-release syndrome and neurotoxicity. Seven CR patients received no further treatment, and 3 of them remained in remission at 6, 6.6, and 14 months after infusion. Eleven CR patients were promptly bridged to transplantation, and 8 of them remained in remission at 4.6 to 13.3 months after transplantation, resulted in 1-year leukemia-free survival rate of 71.6% (95% CI, 44.2–99.0). CD22 antigen loss or mutation was not observed to be associated with relapsed patients. Our study demonstrated that our CD22 CAR T-cells was highly effective in inducing remission in r/r B-ALL patients, and also provided a precious window for subsequent transplantation to achieve durable remission.

INTRODUCTION The phase 1/2 clinical trials of CD19 CAR T-cell that led to US Food and Drug Administration approval excluded patients with central nervous system (CNS) involvement, due to concerns for CAR T-cell related neurotoxicity. The clinical trial (ChiCTR1800014457) examined efficacy and safety of sequential different B cell antigen-targeted CAR T-cell therapy for pediatric refractory/relapsed (r/r) Burkitt Lymphoma with secondary CNS involvement. METHODS Twenty-three patients up to 18 years of age with r/r Burkitt Lymphoma were enrolled between January 17, 2018 and November 8, 2019 including 10 patients with secondary CNS involvement. All enrolled patients received the first mCD19 CAR T-cell infusion. The patients who achieved an ongoing complete response (CR) did not receive further therapy; for patients who retained a partial response (PR) or no response (NR) until mCD19 CAR T cells became undetectable in PB by flow cytometry (FCM), the second hCD22 CAR T-cell infusion was initiated so that it did not prematurely interfere with the antitumor effect of mCD19 CAR T cells, and could prevent disease progression in the setting of continuous mCD19 CAR T-cell contraction; and for patients who had progressive disease (PD) or relapsed disease (RD) after responding to mCD19 CAR T cells, considering that the tumor had already produced immune escape from mCD19 CAR T cells, regardless of whether there were mCD19 CAR T cells detectable in PB by FCM, the second hCD22 CAR T-cell infusion was immediately initiated. After the second hCD22 CAR T-cell infusion, the patients who retained a PR until hCD22 CAR T cells became undetectable in PB by FCM received the third hCD20 CAR T-cell infusion; for patients who suffered relapses after achieving the first CR (CR1) following the first infusion and then attained the second CR (CR2) by the second infusion, the third hCD20 CAR T-cell infusion was given as consolidation therapy after loss of hCD22 CAR T cells detectable in PB by FCM, and the patients who had PD after having obtained a PR immediately received the third hCD20 CAR T-cell infusion even if hCD22 T cells were still detectable in PB by FCM. After the third hCD20 CAR T-cell infusion, the patients who retained a PR until hCD20 CAR T cells became undetectable in PB by FCM were given the fourth hCD19 CAR T-cell infusion. RESULTS The median time from the last infusion to cutoff date was 17 months (range, 15 to 23). At 18 months after the last infusion, all patients had an estimated PFS rate of 78% (95% CI, 55 to 90) and OS rate of 83% (95% CI, 60 to 93). Through mCD19 CAR T-cell expansion in the cerebrospinal fluids (CSF) of ten patients with CNS involvement, 8 patients achieved a CR, and 2 PR. Among the 8 patients who achieved a CR, 4 patients (P5, P15, P16, and P19) maintained an ongoing CR, whereas 4 patients (P7, P11, P17, and P21 ) developed a RD at m6, m2, m3, and m1.5 after infusion. P17 died of rapid intracerebral mass (ICM) progression, which resulted in elevated intracranial pressure (ICP), a contraindication to further CAR T-cell therapy. Two patients (P7 and P21) with CNS RD, P11 with bone marrow RD, and 2 patients (P13 and P20) with PR received the second hCD22 CAR T cells infusion. Among 5 patients experiencing 2 cycles of CAR T-cell infusion, 4 patients (P7, P13, P20, and P21) had hCD22 CAR T-cell expansion in CSF. P13 achieved an ongoing CR1; P20 and P21 died of ICM progression, who did not proceed with the third hCD20 CAR T-cell infusion due to elevated ICP and loss of CD20 in tumor cells, respectively; P7 and P11 achieved a CR2, who underwent the third hCD20 CAR T cells infusion, and then had hCD20 CAR T cells expansion in CSF. P11 achieved an ongoing CR2; P7 had a CNS RD again at m3 after obtaining a CR2 and finally died of rapid disease progression. Six patients achieved ongoing CR after the last infusion. The estimated 18-month PFS rate of 60% (95% CI, 25 to 83) in the patients with CNS involvement was not significantly lower than 92% (95% CI, 56 to 99) in the patients without CNS involvement (p=0.28 ). Grade 3 neurotoxicity occurred in 30% (3/10) of patients with CNS involvement. All adverse events were reversible. Statistically significant difference was not observed in the severity of neurotoxicity between patients with and without CNS involvement (p= 0.62). CONCLUSION Sequential CAR T-cell therapy may result in a durable response and is safe in pediatric r/r Burkitt lymphoma. Patients with secondary CNS involvement may benefit from sequential CAR T-cell therapy. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Introduction: Chimeric antigen receptor T-cell (CART) Therapy has induced high complete remission (CR) in the patients with relapsed/refractory (r/r) B-cell acute lymphoblastic leukemia (B-ALL), but the median event-free survival is only 6.1 months (Park JH. et al. NEJM 2018). To achieve durable leukemia-free survival (LFS), it may helpful to receive allogeneic hematopoietic stem cell transplantation (allo-HSCT) after CR with CART therapy. Objective: In current study, safety and long-term outcomes of allo-HSCT after CART therapy in r/r B-ALL were assessed and risk factors for overall survival (OS) and LFS were analyzed. Methods: Between June 2017 to June 2020, 137 patients with r/r B-ALL who achieved CR with CD19 or CD22 CART then bridge to allo-HSCT in our hospital were evaluated. The median age was 9.7 years (1.8-56). Bone marrow (BM) samples were collected to detect somatic gene mutations of leukemic cells. Blood samples from patients, parents, and potential related donors were collected to analyze germline mutations of hematological and immunological hereditary predisposition genes. The disease status before transplant was minimal residual disease (MRD) negative in 114 (83.2%) patients and MRD positive in 23 (16.8%) patients. The median time from CART infusion to allo-HSCT was 51 days (29-94). The types of transplant were haploidentical (105, 76.6%), unrelated (26, 19.0%), and identical siblings (6, 4.4%). Myeloablative reduced-toxicity conditioning (RTC) with either total body irradiation (TBI)/fludarabine-based (84, 61.3%) or busulfan (BU) /fludarabine-based (53, 38.7%) were applied. Cyclosporine, mycophenolate mofetil and short-term methotrexate were employed for graft-versus-host disease (GVHD) prophylaxis. Results: Full donor engraftment was achieved in all patients. The median time for neutrophil and platelet engraftment was 15 days (9-21) and 13 days (5-33). With a median follow-up of 653 days (481-783), two-year OS was 74.1% (64.4-81.5) and two-year LFS was 64.4% (54.6-72.6). The incidences of grade Ⅱ to Ⅳ and severe acute GVHD (aGVHD) were 27%, 14.5%, respectively. Twenty-five (18.2%) cases developed chronic GVHD (cGVHD) (limited 18, extensive 7). The incidences of CMV and EBV viremia were 20.1%, 12.5%. Hemorrhagic cystitis was found in 32 (22.2%) cases. Thirty-six (26.3%) cases relapsed after transplant, 28/114 (24.6%) in MRD negative cohort and 8/23 (43.5%) in MRD positive cohort. Thirty (21.9%) patients died (relapsed 22, infection 5, multiple organ failure 2, thrombotic microangiopathy 1). Transplant-related mortality was only 5.8%. Total 137 patients with 99 kinds of somatic mutations, the most frequent 15 mutations were TP53, CYP2C19, KRAS, NRAS, PTPN11, CREBBP, KMT2D, KIT, NR3C1, STK11, FLT3, NF1, NUDT15, ABL1, ADAMTS13. LFS was significantly reduced in patients with TP53, KIT, NF1 mutation (P < 0.0001; P < 0.025; P < 0.003). Total 114 evaluable patients with 93 kinds of hematological and immunological hereditary predisposition gene mutations, the most frequent 15 mutations were BLTA, SERPINE1, CYP2C19, F7, TP53, DPYD, MLH1, TNFAIP3, YARS2, HIFIA, ATM, EP300, KIT, ADAMTS13, BRCA2. LFS was significantly reduced in patients with germline EP300 mutation (P < 0.00027). Univariate analysis showed that BU-based regimen (P=0.0194), somatic TP53 mutation (P= 0.002), fungal infection (P= 0.0217), viral infection (P=0.0187) and MRD positive before transplant (P= 0.0227) were adverse factors of OS. The negative factors of LFS were BU-based regimen (P=0.0139), germline EP300 mutation (P Conclusions: Our large series and long-term outcomes have demonstrated that LFS and OS have been improved remarkably in r/r B-ALL who achieved CR with CART then quick bridge to RTC allo-HSCT. Our study has shown the somatic and germline mutation profiles in r/r/ B-ALL and found that somatic TP53 mutation and germline EP300 mutation are negative factors for LFS. TBI-based regimen , MRD negative before transplant and aGVHD have positive impact on LFS, and fungal infection, somatic TP53 and germline EP300 have negative impact on LFS. Disclosures No relevant conflicts of interest to declare.

Background: Epstein-Barr virus (EBV) positive hematological malignancies and EBV-associated hemophagocytic lymphohistiocytosis (HLH) are serious illness that is related with chronic active EBV infection (CAEBV). Recently, more and more immune-related gene mutations have been identified to associate with the EBV positive hematological diseases. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a curative therapy for above diseases. Aim: In present clinical study, the germline gene mutations associated with EBV positive hematological malignancies and EBV positive HLH were analyzed and the outcomes of allo-HSCT in those diseases were investigated. Methods: Between January 2018 and June 2021, 7 patients with EBV positive hematological diseases (lymphoma 3, aggressive NK-cell leukemia (ANKL) 1, secondary HLH 3) who received allo-HSCT in our hospital were included. Blood samples from patients, parents and potential related donors were collected before transplantation and whole exome sequencing of the blood DNA samples was performed to detect and analyze their relevance with the clinical features before and after allo-HSCT. The median age was 23 (5-31) years old. The median disease course before transplant was 6 (3-38) months. Before allo-HSCT, 3 patients (HLH 1, lymphoma 1, ANKL 1) were in non-remission (NR), 2 patients with lymphoma were in partial remission (PR) and the other two patients with HLH were in complete remission (CR). Five patients received allo-HSCT from haploidentical donors and 2 patients from unrelated donors (HLA 10/10, 9/10 matched). Myeloablative conditioning regimen with busulfan (0.8mg/kg iv q6h, 3 days)/etoposide (200mg/m 2, 3 days)/fludarabine (30mg/m 2, 5 days) /rabbit anti-T-cell globulin was used. Cyclosporine, mycophenolate mofetil and short-term methotrexate were employed for graft-versus-host disease (GVHD) prophylaxis. Results: All patients achieved durable engraftment. Plasma EBV-DNA was positive in three patients (1200, 880, 84000 copies/ml, respectively) before transplant and has been negative in all patients after transplant. One patient had gastrointestinal bleeding caused by EBV enteritis. Two patients had CMV viremia. Three patients developed grade III acute GVHD and two had extensive chronic GVHD. All GVHD was resolved with immune-suppressants. With the median follow-up time of 21 (7-36) months, all patients are alive and free of their diseases. Seven disease-related gene mutations were detected including FANCA p.S1088F, TGFBR2 p.T315M, IRF7 p.K446R, PRF1 p.R4C, MPEG1 p.P316S, STX11 p.R49Q, and UNC13D p.G863D. The first 5 gene mutations were detected in the patients with lymphoma and ANKL, and the last 2 gene mutations were detected in the patients with HLH. FANCA is associated with bone marrow failure and the other genes are associated with primary immunodeficiency, all of these gene mutations have been reported and affect in part the function of the genes. In particular, STX11 p.R49Q and UNC13D p.G863D have also been reported in HLH patients and the NK cell activity of these patients is lower than normal levels. Apart from the defined gene mutations in primary HLH, recently, many new immunodeficiency gene mutations have been reported to associate with EBV positive malignancies and HLH which suggests immunodeficiency might play a role in the etiology of the diseases. The gene mutations we reported here might be the main gene mutations associated with disease susceptibility and infection risk. Conclusion: Our preliminary results have shown the gene mutation profile related to EBV positive hematological malignancies and HLH. With our transplant protocol, durable reconstitution of hematopoiesis and immune from healthy donors has been achieved successfully which has resulted in 100% disease-free survival and no transplant-related death. Allo-HSCT is a safe and effective curative modality for EBV positive hematological malignancies and HLH. Disclosures No relevant conflicts of interest to declare.

Acute graft-versus-host disease (aGVHD) is one of major complications of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Genetic polymorphisms of interleukin-7 receptor (IL-7R) have an impact on transplant-related mortality (Z Shamim et al. BMT 2006). Our previous studies indicated that hematological and immunological hereditary predisposition genes have potential influence on major complications of allo-HSCT in patients with hematological malignancies (unpublished data). In present study, the influence of hematological and immunological hereditary predisposition genes on aGVHD after allo-HSCT in the patients with hematological malignancies was investigated. Between February 2018 and August 2019, 92 patients with hematological malignancies who underwent allo-HSCT in Beijing Boren Hospital were enrolled. The median age was 17 (1 to 45) years old. The diagnoses included acute myeloid leukemia (37, 40.2%), acute lymphoblastic leukemia (36, 39.1%), non-Hodgkin's lymphoma (13, 14.1%), and myelodysplastic syndrome (6, 6.5%). The disease status before transplant was complete remission in 54 (58.7%), non-remission in 23 (25%), and relapse in 15 (16.3%). Before transplants, blood samples were collected from the patients, their parents and potential related donors to detect hematological and immunological hereditary predisposition genes with whole exon sequencing and validation by sanger sequencing. The donors were from identical siblings (4, 4.3%) or matched unrelated volunteers (8, 8.7%) or haploidentical family members (80, 86.9%). Myeloablative conditioning regimens with either total body irradiation/fludarabine-based or busulfan/fludarabine-based were applied. Anti-thymocyte globulin was used in haploidentical and unrelated transplants. GVHD prophylaxis was with cyclosporine, short-term methotrexate and mycophenolate mofetil. Methylprednisolong 1-2mg/kg daily was used for aGVHD treatment and basiliximab was administrated in severe aGVHD. Forty-eight patients (52.2%) developed aGVHD as aGVHD group and 44 patients (47.8%) have no aGVHD as non-aGVHD group. IL7R gene heterozygous mutations in recipients significantly increased overall aGVHD and also severe aGVHD after allo-HSCT. IL7R gene heterozygous mutations were found in 7/48 patients in aGVHD group, but 0/44 in non-GVHD group (P = 0.041). The incidence of grade III-IV aGVHD in patients with IL7R gene heterozygous mutations was significantly higher than that without IL7R gene heterozygous mutations. (5/7 vs. 7/41, P=0.007). Among the patients with IL7R gene heterozygous mutations in aGVHD group, 5/7 patients (71.4%) had grade III-IV aGVHD. Three of them with grade III aGVHD had the same heterozygous mutation site: c.c1241t; p.t414m and rest 2 patients with grade IV aGVHD had the same heterozygous mutation site: c.g314a; p. S105n. Our preliminary results have shown that IL7R gene heterozygous mutations in recipients have significantly increased the risk of overall aGVHD and severe aGVHD after allo-HSCT in the patients with hematological malignancies. Larger sample size is needed to address this issue. Disclosures No relevant conflicts of interest to declare.

Chromosome abnormalities and gene variants are important factors in prognosis of the patients with hematological malignancies. Our previous study showed that some germline gene mutations-related to hematological and immunological disorders may have negative impact on the complications of allogeneic hematopoietic stem cell transplantation (allo-HSCT). In current study, the influence of both hematological and immunological hereditary predisposition gene variants and tumor gene variants on the outcomes of allo-HSCT in patients with hematological malignancies was studied. Between January 2018 and June 2020, 164 patients with hematological malignancies who underwent allo-HSCT in our hospital were analyzed. The median age was 14 (1 to 67) years old. The diagnosis included acute myeloid leukemia (n=68, 41.5%), acute lymphoblastic leukemia (n=67, 40.8%) and non-Hodgkin's lymphoma (n=29, 17.7%). The disease status before transplant was CR1 in 49 cases (29.9%), CR2 in 63 cases (38.4%), PR in 22 cases (13.4%), and NR in 30 cases (18.3%). Donors were from haploidentical family members (n=125, 76.2%) or identical siblings (n=18, 11.0%) or unrelated volunteers (n=21, 12.8%). Myeloablative conditioning regimens with either total body irradiation/fludarabine-based or busulfan/fludarabine-based were applied. Anti-thymocyte globulin was used in haploidentical and unrelated transplants. Graft-versus-host disease (GVHD) prophylaxis was with cyclosporine, short-term methotrexate and mycophenolate mofetil. Before transplant, blood samples from patients, their parents and potential related donors were collected to analyze for more than 700 kinds of hematological and immunological hereditary predisposition genes with whole exon sequencing and validation by sanger sequencing. The average sequencing depth was 150×. At diagnosis or relapse, bone marrow samples from patients were obtained to detect for 339 kinds of tumor genes by Illumina sequencing. The average sequencing depth was more than 3000×. With the median follow-up 12.6 (11.2 to 17.0) months, 105 patients (64.0%) achieved durable remission after allo-HSCT. Forty-seven patients (28.7%) relapsed. Twelve patients (7.3%) died. Total 191 immunodeficiency-related hereditary predisposition gene variants were identified in this cohort (average 3.5 gene variants per patient; range, 0-10). Twenty-six of them were recurrent more than 6 times that including TYK2, IFIH1, CFTR, LRBA, IL7R, POLE, RNF31, NLRP12, TTC7A, ATM, CARD14, CHD7, NOD2, TNFRSF13B, BLB,CFB, EPG5, C8A, C8B, CFH, IRF, MSH6, NCF2, NFAT5, PMS2 and ST1M1.IFIH1and IL10RB gene variants were poor factors for relapse post-transplant (P=0.006, P=0.007). The functions of these genes involve in combined immunodeficiency, autoinflammatory disease, complement deficiency, immune deficiency, T-cell dysfunction and antibody deficiency. Total 153 tumor gene variants were identified in this patient series (average 1.93 gene variants per patient; range, 0-16). Seventeen of them were recurrent more than 4 times that including TP53, TPN11, KIT, FLT3, NRAS, NUDT15, STK11, CREBBP, NPM1, DNAH9, DNMT3A, IDH2, HEK2, KRAS, KMT2C, NOTCH1 and NR3C1. TP53, KRAS and NUDT15 gene variants were poor factors for relapse after allo-HSCT (P=0.000025, P=0.0082, P=0.000018). Hemophagocytic lymphohistiocytosis (HLH)-related genes variants were found in 9 patients (5.5%). Frequent HLH-related gene variants were in CD27, PRF1, STX11, and UNC13D. Fanconi anemia (FA)-related gene variants were seen in 11 patients (6.7%). Common FA-related gene variants were in BRCA1, BRCA2, BRIP1, FANCA, FANCC, FANCF, FANCM and FANCG. Significantly higher incidences of acute GVHD (aGVHD) and/or infections were noted in the patients with HLH and/or FA-related gene variants compared with those without HLH and FA-related gene variants (p=0.019). Our results have shown that both inherited and acquired hematological and immunological-related gene variant profiles in the patients with hematological malignancies and some recurrent gene variants (IFIH1 and IL10RB; TP53, KRAS and NUDT15) have negative impact on the outcomes of allo-HSCT including leukemia-free survival, aGVHD and infections. Keywords: allogeneic hematopoietic stem cell transplant, gene expression, relapse, hematological malignancy Disclosures No relevant conflicts of interest to declare.

Despite worldwide promising clinical outcome of CD19 CAR-T therapy, relapse after this therapy is associated with poor prognosis and has become an urgent problem to be solved. We conducted a CD22 CAR T-cell therapy in 34 relapsed or refractory (r/r) B-ALL pediatric and adult patients who failed from previous CD19 CAR T-cell therapy. Complete remission (CR) or CR with incomplete count recovery (CRi) was achieved in 24 of 30 patients (80%) that could be evaluated on day 30 after infusion, which accounted for 70.5% of all 34 enrolled patients. Most patients only experienced mild cytokine-release syndrome and neurotoxicity. Seven CR patients received no further treatment, and 3 of them remained in remission at 6, 6.6, and 14 months after infusion. Eleven CR patients were promptly bridged to transplantation, and 8 of them remained in remission at 4.6 to 13.3 months after transplantation, resulted in 1-year leukemia-free survival rate of 71.6% (95% CI, 44.2-99.0). CD22 antigen loss or mutation was not observed to be associated with relapsed patients. Our study demonstrated that our CD22 CAR T-cells was highly effective in inducing remission in r/r B-ALL patients, and also provided a precious window for subsequent transplantation to achieve durable remission.

The clinical utility of Traditional Chinese Medicine (TCM) herbs/roots extracts in osteoporosis (OP) and osteoarthritis (OA) has been described in multiple reports, but there have been few studies of TCM for preventing bone loss and cartilage degradation simultaneously. Six-month-old female Sprague-Dawley rats each were subjected to ovariectomized (OVX) or sham surgery and treated orally once daily with herbal extracts or vehicle. Body weight was recorded weekly, and blood samples were collected from fasting animals at different time points. Biochemical markers of bone resorption and cartilage degradation were analyzed. Changes in bone mineral density and calcium content were determined in the femoral center and femoral telocentric end of rats. Out of 56 TCM herbs/roots extracts, only kudzu root demonstrated consistent joint protective effects. OVX resulted in a marked increase in bone resorption and cartilage degradation, which could be significantly reversed by kudzu after three weeks of treatment. Compared to vehicle, kudzu induced a significant increase in bone mineral density in the femoral center and femoral telocentric end, and calcium content. The results show that kudzu exerts direct effects on articular cartilage in the OVX rat and can effectively prevent the acceleration of cartilage degradation induced by ovariectomy. Moreover, kudzu has demonstrated positive effects on metabolic health (cause a weight reduction) and may represent a possible treatment for OP and OA with high body mass index. Further studies are needed to investigate the potential effects of kudzu root in postmenopausal women.

The clinical utility of Traditional Chinese Medicine (TCM) herbs/roots extracts in osteoporosis (OP) and osteoarthritis (OA) has been described in multiple reports, but there have been few studies of TCM for preventing bone loss and cartilage degradation simultaneously. Six-month-old female Sprague-Dawley rats each were subjected to ovariectomized (OVX) or sham surgery and treated orally once daily with herbal extracts or vehicle. Body weight was recorded weekly, and blood samples were collected from fasting animals at different time points. Biochemical markers of bone resorption and cartilage degradation were analyzed. Changes in bone mineral density and calcium content were determined in the femoral center and femoral telocentric end of rats. Out of 56 TCM herbs/roots extracts, only kudzu root demonstrated consistent joint protective effects. OVX resulted in a marked increase in bone resorption and cartilage degradation, which could be significantly reversed by kudzu after three weeks of treatment. Compared to vehicle, kudzu induced a significant increase in bone mineral density in the femoral center and femoral telocentric end, and calcium content. The results show that kudzu exerts direct effects on articular cartilage in the OVX rat and can effectively prevent the acceleration of cartilage degradation induced by ovariectomy. Moreover, kudzu has demonstrated positive effects on metabolic health (cause a weight reduction) and may represent a possible treatment for OP and OA with high body mass index. Further studies are needed to investigate the potential effects of kudzu root in postmenopausal women.

Introduction: The development of hematological malignancies may be relevant to hereditary predisposition and harmful external environment. Germ line predisposition to myeloid neoplasms has been incorporated in the WHO 2016 classification of myeloid neoplasms. Hereditary predisposition genes may also play a certain role in the complications of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Objective: In present clinical study, we investigate whether hematological and immunological hereditary predisposition genes have influence on the outcomes of allo-HSCT in patients with hematological malignancies. Methods: Between January 2018 and May 2019, ninety-one patients with hematological malignancies who underwent allo-HSCT with the same protocols in our hospital were enrolled. The median age was 20 (2 to 66) years old. Male to female was 1.53:1. The diagnosis included AML (n=32, 35.2%), ALL (n=47, 51.6%) and NHL (n=12, 13.2%). The disease status before transplants was CR1 in 31 cases (34.1%), CR2 in 45 cases (49.5%), PR in 5 cases (5.5%), and NR in 10 cases (11.0%). Hematological and immunological hereditary predisposition genes of the patients, their parents and potential related donors were prospectively detected before transplant with whole exon sequencing and validation by sanger sequencing. Donors were from haploidentical family members (n=74, 81.3%) or identical siblings (n=6, 6.6%) or unrelated volunteers (n=11, 12.1%). Myeloablative conditioning regimens with either TBI/Fludarabine or Busulfan/Fludarabine were applied. Anti-thymocyte globulin (ATG) was used in haploidentical and unrelated transplants. Graft-versus-host disease (GVHD) prophylaxis was with cyclosporine, short-term methotrexate and mycophenolate mofetil. Prevention of fungal, pneumocystis carinii and herpes virus infections was routinely administrated. Chi-square test was used to analyze whether hereditary predisposition genes were associated with major complications after allo-HSCT. The study used the IBM SPSS Statistics software for analysis. P value Results: Thirty-one (34.1%) cases carried primary hemophagocytic lymphohistiocysis (HLH) related gene variants and the average number of gene variants was 1.2 (range 1 to 2). More frequent HLH gene variants were in UNC13D, LYST, PRF1, AP3B1 and STX11. Forty-four (48.4%) cases carried Fanconi anemia (FA) related gene variants and the average number of gene variants was 1.27 (range 1 to 3). More frequent FA gene variants were in BRCA2, FANCA, PALB2, FANCD2, BRCA1, FANCL, FANCI and SLX4. Ninety (98.9%) cases carried immunodeficiency related gene variants and the average number of gene variants was 3.25 (range 1 to 10). More frequent immunodeficiency gene variants were in IFIH1, CHD7, TYK2, LRBA, NFAT5, IL7R, NLRP12, POLE, TNFRSF13B, ATM, IGLL1, ORAI1, RNF31, C8A, CBLB, IRF8, LIG1, NCF2, NCF4, STIM1 and TCF3 .The incidence of acute GVHD (aGVHD) in patients carried HLH gene variants was significantly higher than that without HLH gene variants (45.1% vs. 21.6%, p=0.020). Bacterial infections occurred more frequent in patients with FA gene variants than that without FA gene variants (47.7% vs. 21.3%, p = 0.008). There was a trend of higher relapse rate in patients with more than 3 immunodeficiency gene variants than that with less than 3 immunodeficiency gene variants but without statistical significance (23.8% vs. 11.4%, p > 0.05). Conclusions: Our preliminary results have shown that hematological and immunological hereditary predisposition genes have potential influence on major complications of allo-HSCT in patients with hematological malignancies. HLH gene variants increase the risk of aGVHD, and FA gene variants increase bacterial infections after allo-HSCT. The patients with multiple immunodeficiency related gene variants may increase relapse rate post-transplant. A larger cohort and longer follow-up are needed to address this issue. Disclosures No relevant conflicts of interest to declare.

ObjectiveThe specific degradation of type II collagen and aggrecan by matrix metalloproteinase (MMP)-9, -13 and ADAMTS-4 and -5 (aggrecanase-1 and -2) in the cartilage matrix is a critical step in pathology of osteoarthritis (OA). The aims of this study were: i) To investigate the relative contribution of ADAMTS-4 and ADAMTS-5 to cartilage degradation upon catabolic stimulation; ii) To investigate the effect of regulating the activities of key enzymes by mean of broad-spectrum inhibitors.MethodsBovine full-depth cartilage explants stimulated with tumor necrosis factor alpha (TNF-α) and Oncostatin M (OSM) were cultured for 21 days with or without a number of inhibitors targeting different types of proteases. Monoclonal antibodies were raised against the active sites of ADAMTS-4, -5, MMP-9 and -13, and 4 ELISAs were developed and technically validated. In addition, the established AGNxI (ADAMTS-degraded aggrecan), AGNxII (MMP-degraded aggrecan), and CTX-II (MMP-derived type II collagen) were quantified in the explants-conditioned media.ResultsWe found that: i) Active ADAMTS-4, MMP-9, -13 were released in the late stage of TNF-α/ OSM stimulation, whereas no significant active ADAMTS-5 was detected in either extracts or supernatants; ii) Active ADAMTS-4 was primarily responsible for E373-374A bond cleavage in aggrecan in this setting; and iii) The compensatory mechanism could be triggered following the blockage of the enzyme caused by inhibitors.ConclusionsADAMTS-4 appeared to be the major protease for the generation of 374ARGS aggrecan fragment in the TNF-α/OSM stimulated bovine cartilage explants. This study addresses the need to determine the roles of ADAMTS-4 and ADAMTS-5 in human articular degradation in OA and hence identify the attractive target for slowing down human cartilage breakdown.