The adverse event after the CART-30 cell infusion.

Supplementary Table S1. Phonotypical characteristics of PBMCs and expanded T Cells as well as transduction efficiency of infused cell products. Supplementary Table S2. Detailed prior chemotherapy regimens for patients before CD30-CART treatment. Supplementary Table S3. Statistics of patient characteristics. Supplementary Table S4. The absolute doses of conditioning regimens for all enrolled patients. Supplementary Table S5. Toxicities possibly related to CD30-CART infusion-containing protocol. Supplementary Table S6. Maximum change from baseline of target measurable lesions. Supplementary Table S7. Clinical response in CD30-CAR T cell- treated patients.

Clinical response of primary-refractory HL in patients receiving CART-30 cell therapy.

Case selection criteria;Methods;Figure Legends

Maximum reduction in target lesion size.

Progression-free survival by Kaplan-Meier method.

Chimeric antigen receptor (CAR)-modified T cells have been reported to successfully treat patients with hematological malignancies. 1,24][5] These challenges may restrict the application of autologous CAR T cells in the clinic.Recent studies have indicated the feasibility of using allogeneic universal CAR T cells to treat infant, pediatric, and adult patients with leukemia. 6,7However, although allogeneic universal CAR T cells that disrupt the T-cell receptor (TCR) a chain to reduce the expression of TCR could reduce the occurrence of graft-versus-host disease (GVHD), these allogeneic cells can be rapidly rejected by the host's immune system because of their expression of human leukocyte antigen (HLA).][10] Reduced alloreactivity of TCR and B2M double-disrupted CAR T cells in vitro and in animal models has been observed. 8,11Therefore, universal CAR T cells with double disruption of TCR and B2M may be a candidate for the treatment of patients with cancer.We generated TCR and B2M double-disrupted universal CAR T cells from healthy donor T cells using lentivirus and CRISPR/Cas9 genome-editing technology, and their in vitro characteristics and specific antitumor efficacy were confirmed as shown in supplemental Figure 1.In addition, we report 2 cases of relapsed/refractory diffuse large B-cell lymphoma (DLBCL) in which patients received universal CAR T-cell therapy.Additional details regarding the study procedures are provided in the data supplement.

Chimeric antigen receptor T (CAR T) cells targeting CD19 have achieved breakthroughs in the treatment of hematological malignancies, such as relapsed/refractory non-Hodgkin lymphoma (r/rNHL); however, high rates of treatment failure and recurrence after CAR T cell therapy are considerable obstacles to overcome. In this study, we designed a series of tandem CARs (TanCARs) and found that TanCAR7 T cells not only showed dual antigen targeting of both CD19 and CD20 but also formed superior and stable immunological synapse (IS) structures, which may be related to their robust antitumor activity. In an open-label, single-arm phase I/IIa trial (ClinicalTrials.gov number NCT03097770), we enrolled 33 patients with r/rNHL, and a total of 28 patients received an infusion after conditioning chemotherapy. The primary objective was to evaluate the safety and tolerability of TanCAR7 T cells. Efficacy, progression-free survival and overall survival were evaluated as secondary objectives. Cytokine release syndrome (CRS) occurred in 14 patients (50%), with 36% grade 1 or 2 and 14% grade 3. No cases of CAR T cell-related encephalopathy syndrome (CRES) of grade 3 or higher were confirmed in any patient. One patient died from a treatment-associated severe pulmonary infection. The overall response rate was 79% (95% confidence interval [CI], 60 to 92), and the complete response rate was 71%. The progression-free survival rate at 12 months was 64% (95% CI, 43 to 79). In this study, TanCAR7 T cells elicited a potent and durable antitumor response but not grade 3 or higher CRES in patients with r/rNHL.

An amendment to this paper has been published and can be accessed via the original article.

Abstract Purpose: Patients with relapsed/refractory primary mediastinal B-cell lymphoma (rrPMBCL) represent a particularly challenging population to treat, with few life-saving treatment options in the context of a dismal prognosis. Patients and Methods: In this open-label, single-arm, phase II study, the safety and efficacy of combined regimen of chemotherapy consisting of gemcitabine, vinorelbine, and pegylated liposomal doxorubicin (GVD) plus anti-PD-1 antibody camrelizumab was assessed in rrPMBCL. Patients received chemo-immunotherapy every 3 weeks until the second confirmed complete response (CR) or up to 12 cycles, followed by camrelizumab monotherapy for up to 1 year. The primary endpoints were objective response rate (ORR) and safety. Results: Twenty-seven response evaluable patients were enrolled, who received a median of three first-line therapies, 59% with bulky disease. The ORR was 74%, including 56% with a CR. A median time of 1.7 months to response was observed, with 78% exhibiting tumor shrinkage at the first evaluation. After 24.8 months median follow-up, the median duration of response was not reached, with a 65% 2-year estimated response rate. Thirteen responders remained in sustained complete remission. Estimated 24-month progression-free survival and overall survival rates were 48.2% and 81.5%, respectively. Any grade and grade 3 treatment-related adverse events (AE) occurred in 93% and 33% of patients, respectively; with no grade 4 or 5 AEs. Baseline levels of IL10, IFNγ, and soluble Fas were associated with objective response. Conclusions: Camrelizumab plus GVD chemotherapy offers a potent option as life-saving chemo-immunotherapy with promising efficacy and a manageable safety profile for patients with rrPMBCL, especially with bulky aggressive disease.

Introduction Since primary mediastinal B-cell lymphoma (PMBCL) is a rare cancer, a prospective clinical trial is challenging and optimal therapy for relapsed/refractory PMBCL (rrPMBCL) has not been defined. The current clinical standard-of-care has been extrapolated from retrospective series or from subgroup analyses of prospective trials designed for diffuse large B-cell lymphoma. Novel salvage strategies are therefore urgently required in rrPMBCL. Methods This single-arm, open-label, phase II trial enrolled patients with heavily pre-treated, rrPMLBCL who have bulky disease (the longest diameter ≥5 cm) and have relapsed or refractory disease after ≥ 2 lines of prior therapy. All enrolled patients received combining PD-1 blockade using Camrelizumab with GVD chemotherapy (gemcitabine, vinorelbine and pegylated liposomal doxorubicin)every 3 weeks until the patients achieved second assessable complete response or up to 12 cycles, followed by maintenance camrelizumab monotherapy up to 1 year. Safety was assessed by CTCAE v5.0 in the population consisted of all patients who received ≥ 1 dose of study drug. Clinical response was evaluated by computed tomography (CT) every 2 cycles and by positron emission tomography-computed tomography (PET-CT) every 4 cycles according to the Lugano Response Criteria for Non-Hodgkin's Lymphoma. Peripheral blood samples were collected before every cycle and tumors were biopsied prior to initiation of therapy for relevant biomarker analysis. The primary end points were objective response rate (ORR) and safety/tolerability. Key secondary end points were complete response rate (CRR), progression-free survival (PFS) and overall survival (OS). Results At the analysis cutoff date (March 31, 2019), 27 rrPMLBCL patients were enrolled: median age 30 years (range: 18-45), 52% female (14/27), median 3 lines of prior therapy (range: 2-6), median 6 cycles of rituximab-containing regimens (range: 2-10). In all 27 response-evaluable patients with heavily pretreated rrPMBCL, Camrelizumab plus GVD achieved unprecedented OR (74.1% [95% CI 55.3-86.8]) and CR rates (55.6% [95% CI 37.3-72.4]), with rapid and lasting responses. The median time to response of 20 patients achieving response was 1.7 months (95% CI 1.6-1.8), while 21 (78.0% [95% CI 59.2-89.4]) patients had a decrease in target lesions at the first tumor scan assessment. With a median follow-up of 12.5 months (range, 3.2-20.6), the median duration of response (DOR) and progression-free survival (PFS) were not reached. Censored at 1-year, the DOR and PFS was 80.0% (95% CI 55.1-92·0) and 59·0% (95% CI 38·3-74·9), respectively. Sixteen of 20 (80%) responders, including all 15 patients achieving complete response, had an ongoing response at data cutoff. The addition of Camrelizumab to the GVD regimen did not appear to exacerbate treatment-related adverse events. Any-grade and grade 3 treatment-related adverse events was occurred in 25 (93%) and 9 (33%) patients, respectively. No grade 4 or 5 adverse events were reported. The most common treatment-related adverse events were neutropenia (70%), leukocytopenia (52%), pruritus (30%) and anaemia (26%). An exploratory analysis undertaken to identify potential biomarkers of response indicated a role for serum levels of IFN-γ, IL-10 and sFas to predict the clinical outcome. Conclusions Patients with rrPMBCL represent a particularly challenging population to treat, with few effective treatment options in the context of a dismal prognosis. Aside from palliation, new salvage therapeutic strategies are urgently needed. Camrelizumab plus GVD chemotherapy should be considered as a compelling new standard of salvage therapy with promising efficacy and manageable safety profile for patients with rrPMBCL. Figure Disclosures Rasko: Cure The Future Foundation: Membership on an entity's Board of Directors or advisory committees; Genea: Equity Ownership; Takeda: Honoraria; Rarecyte: Consultancy, Equity Ownership; Gene Technology Technical Advisory, Australian Government: Other: Advisory committee; GSK: Honoraria.

7556 Background: Relapsed/refractory PMBCL (rrPMBCL) generally has limited treatment options and dismal prognosis. Recently, monotherapy of anti-PD-1 was reported to induce an effective and durable clinical response in nearly half PMBCL patients. However, the use of anti-PD-1 alone is notably unpractical for those with bulky aggressive lesions or life-threatening tumor mass given that anti-PD-1 alone always induce delayed tumor degradation. This ongoing, phase I/II study was aimed to evaluate the safety and efficacy of GVD chemo-regimen plus anti-PD-1 (SHR-1210) with or without low-dose decitabine priming in rrPMBCL patients with bulky aggressive lesions. Methods: This trial is enrolling patients with rrPMBCL who have bulky disease (minimum measurement must be > 75 mm in the longest diameter) with aggressive phenotype, rapid progression and fatal prediction. Enrolled patients were randomizedly assigned into 2 salvage treatment cohorts: GVD (Gemcitabine 0.8 g/m2,Vinorelbine 30 mg/d, Doxorubicin 20 mg/m2 , day 1, per 3 weeks) plus SHR-1210 (4 mg/kg, day 2, per 3 weeks) with (cohort 1) or without (cohort 2) low-dose decitabine priming (10mg/d, day -1 to -5, per 3 weeks). Safety was assessed by CTCAEv4.0, and clinical response by International Working Group (IWG) Response Criteria. Results: The prominent adverse event is ≤Grade 3 hematotoxicity in cohort 1. All enrolled 18 heavily-pretreated PMBCL patients had an effective control of disease progression after 1-cycle treatment and no death incident occurred so far. 11 patients had completed 4- to 8-cycle treatment, among them, 7 from cohort 1 were 2 CR, 4 PR, and 1 SD; 4 from cohort 2 were 3 CR and 1 PR. 6 patients who finished 8-cycle treatment had a 6- to 10-month ongoing progression-free survival so far. Conclusions: Both 2 regimens had comparably life-saving and durable clinical efficacy. Although all observed adverse events were tolerable in both groups, patients enrolled in the future will by treated by regimen without decitabine in view of the relatively lower treatment-associated toxicities. Clinical trial information: NCT03346642.

Excessive activated T-cell proliferation was observed in vivo in one patient after an anti-CD19-chimeric antigen receptor (CAR) T-cell infusion. The patient, who had chemotherapy refractory and CD19+ diffuse large B-cell lymphoma (DLBCL), received an anti-CD19 CAR T-cell infusion following conditioning chemotherapy (fludarabine/cyclophosphamide). The lymphocyte count in the peripheral blood (PB) increased to 77 × 109/L on day 13 post infusion, and the proportion of CD8+ actived T cells was 93.06% of the lymphocytes. Then, the patient suffered from fever and hypoxaemia. Significant increases in serum cytokine, lactate dehydrogenase, aspartate aminotransferase (AST), alanine transaminase (ALT), and glutamic-oxalacetic transaminase (γ-GT) levels were observed. A high-throughput sequencing analysis for T-cell receptors (TCRs) and whole-genome sequencing were used to explore the mechanisms underlying this excessive T-cell proliferation. TCR diversity was demonstrated, but no special gene mutation was found. The patient was found to be infected with the John Cunningham polyomavirus (JCV). It cannot be ruled out the bystander activation pathway induced by JCV infections related the excessive activated T-cell proliferation. Although the clinical and laboratory data do not fully explain the reason for excessive T-cell proliferation after the anti-CD19 CAR T-cell infusion, the risk of this type of toxicity should be emphasized. This study was registered at www.clinicaltrials.gov as NCT01864889.

目的:探讨靶向CD30的嵌合抗原受体T细胞(CART-30)治疗复发难治性霍奇金淋巴瘤的安全性及有效性,建立相对成熟的治疗体系.方法:2014-2015年在我院对18例复发难治性霍奇金淋巴瘤患者进行CART-30细胞输注治疗,并随访观察.结果:18例患者均顺利接受细胞回输并完成随访,耐受性好,仅有2例患者出现≥3级毒副反应.7例患者疗效评估为部分缓解,6例为疾病稳定,客观反应率为39％(7/18),疾病控制率为72％(13/18),无疾病进展时间平均为9.5个月.结论:CART-30细胞治疗的安全性与有效性结合相对成熟的治疗体系,将使其成为复发难治性霍奇金淋巴瘤的一个新型治疗途径.

e14555 Background: Anti-PD1 have produced significant antitumor activity in multiply malignancies, however, resistance to anti-PD1 is becoming increasingly apparent in recent years. Low-dose decitabine, a classical DNA hypomethylating agent, was shown to boost effector T cell function and trigger a T cell-mediated response. This phase I study was designed to assess the safety, immunological effects and antitumor activity of this novel combination in patients with advancedanduntreated lymphomas and solid tumors. Methods: Patients were treated with decitabine (10mg/d on day 1-5) and anti-PD1 (2-3mg/kg, day 8) per 3 weeks. Modified salvage regimens (lymphoma: COP; solid tumors: platinum-based chemotherapy) were allowed to be intermittently inserted for patients with aggressive progression. Treatment continued unless disease progression or severe toxicity. Safety was assessed by CTCAEv4.0, and response by standard international criteria. The phenotype and activity of T cells were periodically measured in peripheral blood by flow cytometry. Results: Todate, 11 patients with heavily treated history and refractory bulky lesions have been enrolled, including 8 with lymphomas (7 NHL, 1 HL resistant to anti-PD1) and 3 with metastatic solid tumors (2 gastric cancers, 1 esophageal cancer) failure to anti-PD1. 5 patients (45%) experienced Grade ≥ 3 toxicities, with 1 taken off due to toxicity, and 1 died of asystole during the term of severe cytokine release syndrome (CRS). The common events of leukocytopenia and CRS were prominent features of anti-PD1 plus decitabine. 9 patients were evaluable for response, 1 HL obtained complete response, 3 NHL and 3 solid tumors achieved partial response, and 2 NHL had stable disease with nearly 20% shrinkage. The frequency of interferon-γ-producing CD8+ T cells in the total CD3+ population was largely increased after anti-PD1 plus decitabine infusion. Conclusions: Decitabine augmented the pro-inflammatory effects of anti-PD1 characterized by systemic inflammation response, and further improved antitumor activity of anti-PD1. Clinical trial information: NCT02961101. Clinical trial information: NCT02961101.

Abstract Purpose: Relapsed or refractory Hodgkin lymphoma is a challenge for medical oncologists because of poor overall survival. We aimed to assess the feasibility, safety, and efficacy of CD30-targeting CAR T cells in patients with progressive relapsed or refractory Hodgkin lymphoma. Experimental Design: Patients with relapsed or refractory Hodgkin lymphoma received a conditioning chemotherapy followed by the CART-30 cell infusion. The level of CAR transgenes in peripheral blood and biopsied tumor tissues was measured periodically according to an assigned protocol by quantitative PCR (qPCR). Results: Eighteen patients were enrolled; most of whom had a heavy treatment history or multiple tumor lesions and received a mean of 1.56 × 107 CAR-positive T cell per kg (SD, 0.25; range, 1.1–2.1) in total during infusion. CART-30 cell infusion was tolerated, with grade ≥3 toxicities occurring only in two of 18 patients. Of 18 patients, seven achieved partial remission and six achieved stable disease. An inconsistent response of lymphoma was observed: lymph nodes presented a better response than extranodal lesions and the response of lung lesions seemed to be relatively poor. Lymphocyte recovery accompanied by an increase of circulating CAR T cells (peaking between 3 and 9 days after infusion) is a probable indictor of clinical response. Analysis of biopsied tissues by qPCR and immunohistochemistry revealed the trafficking of CAR T cells into the targeted sites and reduction of the expression of CD30 in tumors. Conclusions: CART-30 cell therapy was safe, feasible, and efficient in relapsed or refractory lymphoma and guarantees a large-scale patient recruitment. Clin Cancer Res; 23(5); 1156–66. ©2016 AACR.

Long-term safety and efficacy of CART-20 cells in patients with refractory or relapsed B-cell non-Hodgkin lymphoma: 5-years follow-up results of the phase I and IIa trials

Anti-CD19 chimeric antigen receptor-modified T (CAR-T-19) cells have emerged as a powerful targeted immunotherapy for B-cell lineage acute lymphoblastic leukemia with a remarkable clinical response in recent trials. Nonetheless, few data are available on the subsequent clinical monitoring and treatment of the patients, especially those with disease recurrence after CAR-T-19 cell infusion. Here, we analyzed three patients who survived after our phase I clinical trial and who were studied by means of biomarkers reflecting persistence of CAR-T-19 cells in vivo and predictive factors directing further treatment. One patient achieved 9-week sustained complete remission and subsequently received an allogeneic hematopoietic stem cell transplant. Another patient who showed relapse after 20 weeks without detectable leukemia in the cerebrospinal fluid after CAR-T-19 cell treatment was able to achieve a morphological remission under the influence of stand-alone low-dose chemotherapeutic agents. The third patient gradually developed extensive extramedullary involvement in tissues with scarce immune- cell infiltration during a long period of hematopoietic remission after CAR-T-19 cell therapy. Long-term and discontinuous increases in serum cytokines (mainly interleukin 6 and C-reactive protein) were identified in two patients (Nos. 1 and 6) even though only a low copy number of CAR molecules could be detected in their peripheral blood. This finding was suggestive of persistent functional activity of CAR-T-19 cells. Combined analyses of laboratory biomarkers with their clinical manifestations before and after salvage treatment showed that the persistent immunosurveillance mediated by CAR-T-19 cells would inevitably potentiate the leukemia-killing effectiveness of subsequent chemotherapy in patients who showed relapse after CAR-T-19-induced remission.

Patients with relapsed or refractory non-Hodgkin lymphoma have a dismal prognosis. Chimeric Antigen Receptor (CAR)-modified T cells (CART cells) that targeted CD20 were effective in a phase I clinical trial for patients with advanced B-cell lymphomas. We performed a phase IIa trial to further assess the safety and efficacy of administering autologous anti-CD20 CART (CART-20) cells to patients with refractory or relapsed CD20+ B-cell lymphoma. Eleven patients were enrolled, and seven patients underwent cytoreductive chemotherapy to debulk the tumors and deplete the lymphocytes before receiving T-cell infusions. The overall objective response rate was 9 of 11 (81.8%), with 6 complete remissions (CRs) and 3 partial remissions; no severe toxicity was observed. The median progression-free survival lasted for >6 months, and 1 patient had a 27-month continuous CR. A significant inverse correlation between the levels of the CAR gene and disease recurrence or progression was observed. Clinically, the lesions in special sites, specifically the spleen and testicle, were refractory to CART-20 treatment. Collectively, these results together with our data from phase I strongly demonstrated the feasibility and efficacy of CART-20 treatment in lymphomas and suggest large-scale patient recruitment in a future study. This study was registered at www.clinicaltrials.org as NCT01735604.

Adoptive immunotherapy with T cells expressing a tumor-associated chimeric antigen receptor (CAR) provides a promising approach for tumor therapy. We designed a clinical trial for multiple myeloma (MM) treatment with CAR-modified T cells recognizing CD138 (CART-138). Five patients diagnosed with chemotherapy-refractory MM were enrolled into this trial, although one later advanced to plasma cell leukemia. By intravenous infusions, these patients received CD3+ CART-138 cells in an escalating dose. No intolerable toxicity was observed during this process. CART-138 cells were expanded to a level 1000 times higher than the initial engraftment level and were maintained in the peripheral blood. In addition, increased CART-138 cells were also detected in the bone marrow. Four of the five patients had stable disease (SD) longer than three months, and one patient with advanced plasma cell leukemia had a reduction of the myeloma cells in her peripheral blood (from 10.5% to <3%). This study suggests that the treatment of CART-138 is safe, feasible, and tolerable and has potential antitumor activity in vivo, warranting further research in MM treatment using CART-138.