Early Signals of Anti-Tumor Efficacy and Safety with Autologous CD5.CAR T-Cells in Patients with Refractory/Relapsed T-Cell Lymphoma

Abstract

Introduction We present results of a Phase I clinical trial (NCT03081910) of autologous T-cells expressing a CD5-specific chimeric antigen receptor (CD5.CAR) as therapy for refractory or relapsed (r/r) mature T-cell lymphoma (TCL). T-cells expressing an optimized second-generation, CD28-costimulated CD5.CAR resist fratricide by rapidly degrading CD5 protein and expand normally ex vivo. We evaluated the feasibility of manufacturing autologous CD5.CAR T-cells from patients with r/r peripheral or cutaneous TCL and the safety of these CD5.CAR T-cells as a bridging therapy to allogeneic hematopoietic stem cell transplant (alloHSCT). We also assessed anti-tumor activity and kinetics of in vivo expansion and persistence of CD5 CAR T-cells as well as their off-tumor activity against normal circulating T-cells. Methods and patient characteristics CD5.CAR T-cells were manufactured from PBMC using gammaretroviral transduction of T-cells following anti-CD3/anti-CD28 stimulation. Fourteen patients were enrolled, and we have treated nine patients (age 27-71, median 63) with mature TCL including cutaneous TCL (n=2), angioimmunoblastic TCL (AITL, n=2), peripheral TCL (n=4), and adult T-cell leukemia/lymphoma (n=1). All patients were eligible for alloHSCT and had an available donor. Median number of prior lines of therapy was 5 (2-18), and 5 patients had relapsed after autologous (n=3) or allogeneic HSCT (n=2). Results All patients received autologous CD5.CAR T-cells on one of three dose levels: two on dose level 1 (1x10 7 CAR T cells/m 2), four on dose level 2 (5x10 7 CAR T cells/m 2), and three on dose level 3 (1x10 8 CAR T cells/m 2). Seven patients received a single dose of CD5.CAR T-cells following lymphodepleting chemotherapy (LDC) with cyclophosphamide and fludarabine, and two patients received an additional dose following initial disease evaluation. One patient received a 2 nd course of LDC prior to re-infusion. CD5.CAR T-cells expanded and persisted in all patients for up to 9 months , peaking in peripheral blood at 1-4 weeks post-infusion (Fig. 1). CD5.CAR T-cell infusions were associated with a transient reduction in peripheral T-cell counts but depletion was incomplete as circulating normal T-cells were detected in most patients over the follow-up period. No severe infections were observed. Max grade CRS and ICANS was grade 2, which occurred concurrently in one patient and promptly resolved following treatment. Three patients experienced grade 1 CRS and no other neurotoxicity events were observed. The most frequent treatment related adverse events were cytopenias which recovered to baseline within 28 days in all but 3 patients, in one of whom cytopenias extended beyond day 56. The remaining grade 3 events were primarily non-hematologic laboratory abnormalities which returned to baseline or improved during the monitoring period. Reponses were evaluated 4-6 weeks post-infusion. Four out of nine patients (44%) achieved responses, enabling three to proceed to alloHSCT. Complete responses were obtained in two patients, one with AITL and one with peripheral TCL. Another patient with AITL achieved a mixed response followed by a second infusion of CD5.CAR T-cells and subsequently proceeded to alloHSCT. A very good partial response was achieved in a patient with HTLV-1 driven ATLL (Fig. 2). Two patients remain alive and in CR for 29 months and 24 months respectively. The remaining patients experienced stable disease (n=1) or progressive disease (n=4). Clinical responses were observed on all dose levels. Objective responses did not correlate with cell dose or magnitude of T-cell expansion. All responses were obtained with CD5.CAR T-cell products produced with a shortened manufacturing method in which CAR T-cells were cryopreserved 3-5 days post-transduction instead of undergoing a standard 7-day expansion. Shortened manufacturing resulted in an enrichment of minimally differentiated CCR7+ CD62L+ T-cells (27-fold mean increase in CD4+, 13-fold increase in CD8+; p=0.01) and a 3.5-fold increase in CD27+ CD8+ T-cells (p=0.04) suggesting minimizing duration of ex vivo expansion increases anti-tumor function of CD5.CAR T-cells. Overall, these results show that even in patients with multiply relapsed/resistant TCL, CD5.CAR T-cells can produce anti-tumor responses that are sufficiently durable to enable them to proceed with alloHSCT without inducing clinically significant T-cell aplasia. Figure 1 Figure 1. Disclosures Rouce: Tessa Therapeutics: Research Funding; Pfizer: Consultancy; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees. Hill: Incyte: Membership on an entity's Board of Directors or advisory committees. Grilley: Allovir: Current equity holder in publicly-traded company, Other: Leadership; QB Regulatory Consulting: Other: Ownership, project management support, Research Funding; Marker: Consultancy, Other: Regulatory and project management support. Heslop: Allovir: Current equity holder in publicly-traded company; Gilead: Membership on an entity's Board of Directors or advisory committees; Tessa Therapeutics: Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Membership on an entity's Board of Directors or advisory committees; Kiadis: Membership on an entity's Board of Directors or advisory committees; Marker Therapeutics: Current equity holder in publicly-traded company; Takeda: Membership on an entity's Board of Directors or advisory committees; Fresh Wind Biotherapies: Membership on an entity's Board of Directors or advisory committees; Kuur Therapeutics: Research Funding; GSK: Membership on an entity's Board of Directors or advisory committees. Brenner: Walking Fish Therapeutics: Membership on an entity's Board of Directors or advisory committees; CellGenix GmbH: Membership on an entity's Board of Directors or advisory committees; Marker Therapeutics: Current equity holder in publicly-traded company; Tessa Therapeutics: Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees, Other: Founder; Abintus: Membership on an entity's Board of Directors or advisory committees; Allogene: Membership on an entity's Board of Directors or advisory committees; Bellicum Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees; Bluebird Bio: Membership on an entity's Board of Directors or advisory committees; Athenex: Membership on an entity's Board of Directors or advisory committees; Memgen: Membership on an entity's Board of Directors or advisory committees; Turnstone Biologics: Membership on an entity's Board of Directors or advisory committees; Coya Therapeutics: Membership on an entity's Board of Directors or advisory committees; TScan Therapeutics: Membership on an entity's Board of Directors or advisory committees; Onkimmune: Membership on an entity's Board of Directors or advisory committees; Poseida Therapeutics: Membership on an entity's Board of Directors or advisory committees; Allovir: Current equity holder in publicly-traded company. Mamonkin: Xenetic Biosciences: Consultancy, Membership on an entity's Board of Directors or advisory committees; Allogene Therapeutics: Consultancy, Other: Licensing payments; Beam Therapeutics: Other: Licensing payments; Fate Therapeutics: Other: Licensing payments.

CAR-T Cell therapy in T-cell malignancies: limitations and solutions

“…Grade 1 CRS and grade 2 CRS were observed in three patients and one patient, respectively. No other neurotoxicity events were observed in this clinical trial [40]. Despite these encouraging findings, more trials are needed to verify the safety and efficacy of CD5 CAR-T cell therapy.…”

Chimeric Antigen Receptor Based Cellular Therapy for Treatment Of T-Cell Malignancies

“…Specifically, CAR-Ts expanded for shorter period of time were associated with better responses. Shortening of expansion from 7 to 3-5 also resulted in an enrichment of minimally differentiated CCR7+ CD62L+ T-cells (27-fold mean increase in CD4+, 13-fold increase in CD8+; p=0.01) and a 3.5-fold increase in CD27+ CD8+ T-cells (p=0.04) (108).…”

Current state of CAR-T therapy for T-cell malignancies

“…The latest data from this trial demonstrated that cytopenia was the most common side effect, but patients could recover within a month. 60 Feng et al reported a case with refractory T-cell lymphoma and CNS (Central Nervous System) infiltration. The patient received modified anti-CD5 CAR-T-cells that contained an IL-15/IL-15 sushi complex.…”