Multiancestry analysis of the HLA locus in Alzheimer’s and Parkinson’s diseases uncovers a shared adaptive immune response mediated by HLA-DRB1*04 subtypes

Across multiancestry groups, we analyzed Human Leukocyte Antigen (HLA) associations in over 176,000 individuals with Parkinson’s disease (PD) and Alzheimer’s disease (AD) versus controls. We demonstrate that the two diseases share the same protective association at the HLA locus. HLA-specific fine-mapping showed that hierarchical protective effects of HLA-DRB1*04 subtypes best accounted for the association, strongest with HLA-DRB1*04:04 and HLA-DRB1*04:07, and intermediary with HLA-DRB1*04:01 and HLA-DRB1*04:03. The same signal was associated with decreased neurofibrillary tangles in postmortem brains and was associated with reduced tau levels in cerebrospinal fluid and to a lower extent with increased Aβ42. Protective HLA-DRB1*04 subtypes strongly bound the aggregation-prone tau PHF6 sequence, however only when acetylated at a lysine (K311), a common posttranslational modification central to tau aggregation. An HLA-DRB1*04-mediated adaptive immune response decreases PD and AD risks, potentially by acting against tau, offering the possibility of therapeutic avenues

Genome Editing in Engineered T Cells for Cancer Immunotherapy

Advanced gene transfer technologies and profound immunological insights have enabled substantial increases in the efficacy of anticancer adoptive cellular therapy (ACT). In recent years, the U.S. Food and Drug Administration and European Medicines Agency have approved six engineered T cell therapeutic products, all chimeric antigen receptor-engineered T cells directed against B cell malignancies. Despite encouraging clinical results, engineered T cell therapy is still constrained by challenges, which could be addressed by genome editing. As RNA-guided Clustered Regularly Interspaced Short Palindromic Repeats technology passes its 10-year anniversary, we review emerging applications of genome editing approaches designed to (1) overcome resistance to therapy, including cancer immune evasion mechanisms; (2) avoid unwanted immune reactions related to allogeneic T cell products; (3) increase fitness, expansion capacity, persistence, and potency of engineered T cells, while preserving their safety profile; and (4) improve the ability of therapeutic cells to resist immunosuppressive signals active in the tumor microenvironment. Overall, these innovative approaches should widen the safe and effective use of ACT to larger number of patients affected by cancer

Tracking the fate and origin of clinically relevant adoptively transferred CD8 +

Adoptively transferred tumor-specific cells can mediate tumor regression in cancers refractory to conventional therapy. Autologous polyclonal tumor-specific cytotoxic T cells (CTL) generated from peripheral blood and infused into patients with metastatic melanoma show enhanced persistence, compared to equivalent numbers of more extensively expanded monoclonal CTL, and are associated with complete remissions (CR) in select patients. We applied high-throughput T cell receptor Vβ sequencing (HTTCS) to identify individual clonotypes within CTL products, track them in vivo post-infusion and then deduce the pre-adoptive transfer (endogenous) frequencies of cells ultimately responsible for tumor regression. The summed in vivo post-transfer frequencies of the top 25 HTTCS-defined clonotypes originally detected in the infused CTL population were comparable to enumeration by binding of antigen peptide-HLA multimers, revealing quantitative HTTCS is a reliable, multimer-independent alternative. Surprisingly, the polyclonal CTL products were composed predominantly of clonotypes that were of very low frequency (VLF) in the endogenous samples, often below the limit of HTTCS detection (0.001%). In patients who achieved durable CRs, the composition of transferred CTLs was dominated (57–90%) by cells derived from a single VLF clonotype. Thus, HTTCS now reveals that tumor-specific CTL enabling long-term tumor control originate from endogenous VLF populations that exhibit proliferative/survival advantages. Along with results indicating that naïve cell populations are most likely to contain cells that exist at VLF within the repertoire, our results provide a strong rationale for favoring T cells arising from VLF populations and with early-differentiation phenotypes when selecting subset populations for adoptive transfer

Safety and Efficacy of Fully Human BCMA CAR T Cells in Combination with a Gamma Secretase Inhibitor to Increase BCMA Surface Expression in Patients with Relapsed or Refractory Multiple Myeloma

Abstract Background: Chimeric antigen receptor T cells (CAR T cells) targeting B cell maturation antigen (BCMA) have demonstrated rapid and deep responses among patients with multi-agent refractory multiple myeloma (MM). BCMA is shed from tumor cells mediated through enzymatic cleavage by the gamma secretase complex, and tumor cells with low levels of BCMA could potentially escape CAR T recognition. We showed previously that gamma secretase inhibitors (GSI) increase BCMA surface density on tumor cells, decrease soluble BCMA levels, and enhance efficacy of BCMA CAR T cells in an immunodeficient mouse model. We have completed accrual to a phase 1 first-in-human trial of escalating doses of BCMA targeted CAR T cells in combination with a GSI (JSMD194) for patients with relapsed or refractory multiple myeloma, and herein report results on the 18 patients accrued to this trial. Methods: Eligible patients had relapsed/refractory MM, with ≥ 10% plasma cells in the bone marrow by CD138 IHC, and measurable disease. CD8+ and CD4+ T cells were enriched by To assess the discrete impact of the GSI on plasma cell BCMA expression, patients received a GSI (JSMD194) monotherapy "run-in" consisting of three oral doses (25 mg) administered 48 hours apart over 5 days. A bone marrow sample was obtained on day 5 and BCMA expression was compared to baseline. Following lymphodepleting chemotherapy, BCMA CAR T cells were infused at a starting dose of 5 x 10 7 CAR+ cells, in combination with JSMD194 dosed orally at 25 mg thrice weekly for three weeks, starting on the day of CAR infusion. Results: From June 2018 to March 2021, 18 patients underwent leukapheresis, run-in with JSMD194, and treatment with BCMA CAR T cells. The median age was 65 years, and patients had received a median of 10 prior lines of therapy (range, 4-19). 67% of patients were refractory to lenalidomide, pomalidomide, bortezomib, carfilzomib, and daratumumab, 72% had high-risk cytogenetic features, and 28% had extramedullary disease. 7/18 (39%) had prior BCMA targeted therapy; other BCMA targeted CAR T cell products had previously been administered to 4/18 patients (22%). All 18 treated patients completed the 5-day run-in with JSMD194. After three oral doses of GSI, increased from a median of 610 to 9563 receptors per cell, or a median of 12-fold (range, 0.2-fold to 157-fold; Figure 1). The only patient that did not demonstrate an increase in BCMA ABC after GSI run-in had previously received BCMA targeted therapy and BCMA expression at screening was virtually absent. 5 patients were treated at 5x10 7 CAR+ cells, 3 were treated at 15x10 7 CAR+ cells, 3 were treated at 30x10 7 CAR+ cells, and 7 were treated at 45x10 7 CAR+ cells dose levels. Treatment was consistent with other BCMA CAR T therapy, with manageable toxicities. One patient experienced a DLT. 95% of patients experienced cytokine release syndrome (CRS), mostly grade 1-2 (83%), and 66% of patients experienced ICANS, predominantly grades 1-2. The overall response rate was 89%, with 14 patients achieving ≥ VGPR, and 8 patients achieving CR (including 5 with sCR). Deep responses were observed at all dose levels; including the first patient treated on trial at (dose level 1) who has maintained a stringent CR (sCR) for over 35 months and 3 of 5 patients at dose level 1 had no evidence of progressive disease for >18 months. With a median follow-up of 20 months, the median PFS is 11 months (95% CI, 6 mos to not reached). Amongst patients without prior exposure to BCMA targeted therapy (n=11), the median PFS has not been reached, while amongst those previously exposed to BCMA targeted therapy (n=7), the median PFS was 2 months. Discussion: In this study combining a GSI with BCMA CAR T cells, we have demonstrated that the combination is safe and tolerable. GSI administration routinely increased BCMA surface density on plasma cells. Further, we have observed durable, rapid responses in a heavily pretreated refractory population of MM patients, of whom a significant proportion had prior treatment with BCMA targeted therapy and CAR T therapy. The combination of BCMA CAR T and GSI may augment anti-tumor activity, even when very low doses of BCMA CAR T cells are administered. Figure 1 Figure 1. Disclosures Cowan: Harpoon: Research Funding; Secura Bio: Consultancy; Sanofi Aventis: Consultancy, Research Funding; GSK: Consultancy; Abbvie: Consultancy, Research Funding; Nektar: Research Funding; Cellectar: Consultancy; Bristol Myers Squibb: Research Funding; Janssen: Consultancy, Research Funding. Pont: Lyell Immunopharma: Other: Has equity interest; SpringWorks Therapeutics: Other: Received consulting income; CellPoint B.V.: Current Employment. Sather: Lyell Immunopharma: Current Employment. Turtle: Arsenal Bio: Current holder of stock options in a privately-held company, Other: Scientific Advisory Board; PACT Pharma: Consultancy; Amgen: Consultancy; Eureka Therapeutics: Current holder of stock options in a privately-held company, Other: Scientific Advisory Board; AstraZeneca: Consultancy, Research Funding; Juno Therapeutics/BMS: Patents & Royalties: Right to receive royalties from Fred Hutch for patents licensed to Juno Therapeutics, Research Funding; Myeloid Therapeutics: Current holder of stock options in a privately-held company, Other: Scientific Advisory Board; TCR2 Therapeutics: Research Funding; T-CURX: Other: Scientific Advisory Board; Asher Bio: Consultancy; Allogene: Consultancy; Century Therapeutics: Consultancy, Other: Scientific Advisory Board; Nektar Therapeutics: Consultancy, Research Funding; Precision Biosciences: Current holder of stock options in a privately-held company, Other: Scientific Advisory Board; Caribou Biosciences: Consultancy, Current holder of stock options in a privately-held company, Other: Scientific Advisory Board. Till: Mustang Bio: Consultancy, Patents & Royalties, Research Funding. Libby: GSK: Research Funding; Janssen: Consultancy, Research Funding; BMS: Research Funding; Genentech: Research Funding. Tuazon: BMS: Current Employment. Shadman: Abbvie, Genentech, AstraZeneca, Sound Biologics, Pharmacyclics, Beigene, Bristol Myers Squibb, Morphosys, TG Therapeutics, Innate Pharma, Kite Pharma, Adaptive Biotechnologies, Epizyme, Eli Lilly, Adaptimmune , Mustang Bio and Atara Biotherapeutics: Consultancy; Mustang Bio, Celgene, Bristol Myers Squibb, Pharmacyclics, Gilead, Genentech, Abbvie, TG Therapeutics, Beigene, AstraZeneca, Sunesis, Atara Biotherapeutics, GenMab: Research Funding. Chapuis: Karkinos Therapeutics: Other: Ownership; Lonza: Other: Intellectual Property; Cullian: Other: Intellectual Property; TScan Therapeutics, Inc.: Consultancy, Other: Ownership; SignalOne Bio: Consultancy, Other: Ownership; Bluebird bio: Other: Intellectual Property; Juno therapeutics: Other: Intellectual Property; Adapyive Biotechnologies Corporation: Other: Ownership/Intellectual Property; Pfizer: Other: Intellectual Property; Affini-T: Other: Ownership; Ridgeline: Consultancy; BioNTech: Consultancy. Maloney: MorphoSys: Honoraria; Genentech: Honoraria; Navan Technologies: Honoraria, Other: Stock options; Celgene: Honoraria, Other: Rights to royalties from Fred Hutchinson Cancer Research Center for patents licensed to Juno Therapeutics/Bristol Myers Squibb; Novartis: Honoraria; Kite Pharma: Honoraria, Other: Research funding was paid to my institution, Research Funding; Juno therapeutics: Other: Research funding was paid to my institution, Research Funding; Celgene: Other: Research funding was paid to my institution, Research Funding; Amgen: Honoraria; BMS: Honoraria, Other: Rights to royalties from Fred Hutchinson Cancer Research Center for patents licensed to Juno Therapeutics/Bristol Myers Squibb; Juno Therapeutics: Honoraria, Other: Rights to royalties from Fred Hutchinson Cancer Research Center for patents licensed to Juno Therapeutics/Bristol Myers Squibb; Umoja: Honoraria; Legend Biotech: Honoraria; A2 Biotherapeutics: Honoraria, Other: Stock options; Janssen: Honoraria. Riddell: Lyell Immunopharma: Other. Green: Seagen Inc.: Research Funding; bristol myers squibb: Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding; Cellectar Biosciences: Research Funding; GSK: Membership on an entity's Board of Directors or advisory committees; Janssen Biotech: Membership on an entity's Board of Directors or advisory committees, Research Funding; Juno Therapeutics: Patents & Royalties, Research Funding; Legend Biotech: Consultancy; Neoleukin Therapeutics: Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Membership on an entity's Board of Directors or advisory committees, Research Funding; SpringWorks Therapeutics: Research Funding. OffLabel Disclosure: JSMD194 - an oral gamma secretase inhibito

Third Generation CD20 Targeted CAR T-Cell Therapy (MB-106) for Treatment of Patients with Relapsed/Refractory B-Cell Non-Hodgkin Lymphoma

Background: Chimeric antigen receptor T-cell (CAR-T) therapy is an effective strategy for patients (pts) with B-cell non-Hodgkin lymphomas (B-NHLs). However, long-term remissions are only observed in ~40% of pts with large cell lymphomas (LCL) treated with FDA-approved CD19-targeted products and there is currently no approved CAR-T for other histologies other than mantle cell lymphoma (MCL). CD20 is a proven therapeutic target for B-NHL, supported by previously approved naked and radiolabeled anti-CD20 antibodies and promising results from bispecific antibodies. CD20-targeted CAR-T is another potential adoptive immunotherapy option that could be utilized in combination or in sequence with CD19 CAR-T. We present interim results of our ongoing phase I/II clinical trial investigating safety and efficacy CD20 CAR-T for high-risk B-NHLs (NCT03277729). Methods: MB-106 is a fully human third-generation CD20-targeted CAR-T with both 4-1BB and CD28 costimulatory domains. We use a continuous reassessment method dose escalation design to find the maximally tolerated dose. Lymphodepletion (LD) consists of cyclophosphamide (Cy) ± fludarabine (Flu). CAR-T cells are administered at one of 4 dose levels (DL): DL1: 3.3 x 105, DL2: 1 x106, DL3: 3.3x106, DL4: 1x107 CAR T cells/kg. CAR-T is infused in the outpatient setting except for the first pts of each dose cohort (overnight observation). Pts with CD20+ B-NHL are eligible, including but not limited to LCL, follicular lymphoma (FL)/indolent histologies and MCL. Prior treatment with CD19 CAR-T is permitted. Treatment response is assessed on day 28 using 2014 Lugano criteria. Cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) were graded per 2014 Lee criteria and the ASTCT consensus grading, respectively. Results: Between 2018-2020, 12 pts underwent leukapheresis (LA) and 11 (92%) were treated on the protocol. The first 7 pts (3 FL, 3 MCL, 1 hairy cell variant) were treated using the original cell manufacturing method with separate culturing of CD4+ and CD8+ cells (“original process”). First 4 of those 7 pts (2 treated at DL1 and 2 at DL2) only received Cy for LD and the other 3 (all at DL1) were treated after LD with Cy-Flu. There was no evidence of ICANS in those 7 pts and only 1 pt developed CRS (grade 3 - unexplained elevated alkaline phosphatase (ALP) in the setting of fever). Responses at 4 weeks for pts treated with the “original process” included stable disease (SD) in 4 and progressive disease (PD) in 3 pts. Given the challenges in meeting the protocol requirements for CD4+ and CD8+ specific doses, poor CAR-T expansion, and lack of clinical responses, enrollment was placed on a hold and cell manufacturing process underwent a major revision which included changing to a combined culture of CD4+ and CD8+ cells, among other modifications (“modified process”). Since December 2019 enrollment was reinitiated at DL1 under the “modified process.” Four pts were treated after LD with Cy-Flu at DL1 (2 FL) and dose level 2 (1 FL and 1 MCL) (details in Table 1). Target cell doses were achieved in all 4 patients and treatment was tolerated with no occurrence of CRS (any grade) or ICANS (any grade). Responses were observed at both dose levels and included complete remissions (CRs) in 2 FL pts (1 treated at DL1 and 1 at DL2), partial remission (PR) in a MCL pt treated at DL2 and PD in a FL pt who was treated at DL1. Robust CAR-T expansion was seen in 3 of the 4 patients, with peak blood levels of CAR+ T cells of 161, 5.5, and 401 CAR+ cells/µl, corresponding to 80%, 23%, and 72% of all CD8+ cells. Combining all 11 pts treated under the “original” and “modified” processes, no dose-limiting toxicity (DLT) was observed. Grade ≥ 3 toxicities included: anemia (n=4, 36%), lymphopenia (n=3,27%), febrile neutropenia (n=2, 18%), hypertension (n=1, 9%) , hypotension (n=1, 9%), thromboembolic event (n=1, 9%), neutropenia (n=1, 9%), elevated ALP (n=1, 9%), pneumonia (n=1, 9%), bacteremia (n=1, 9%), hyperglycemia (n=1, 9%), pleural effusion (n=1, 9%) and generalized pain (n=1, 9%). Conclusion: Early results of our ongoing study of CD20 CAR-T for high-risk NHLs suggests an extremely favorable safety profile (no CRS or ICANS of any grades) with the modified manufacturing process. There is also evidence of clinical activity even at low dose levels with observed CRs at DLs 1 and 2. Enrollment at higher DLs is currently ongoing and the data will be updated at the time of presentation. Download : Download high-res image (340KB) Download : Download full-size image Disclosures Shadman: Abbvie, Genentech, Astra Zeneca, Sound Biologics , Pharmacyclics, Verastem, ADC therapeutics, Beigene, Cellectar, BMS, Morphosys and Atara Biotherapeutics: Consultancy; Mustang Bio, Celgene, Pharmacyclics, Gilead, Genentech, Abbvie, TG therapeutics, Beigene, Astra Zeneca, Sunesis, Beigene: Research Funding. Yeung: OBI;Pfizer: Research Funding; Adaptive Biotechnology;Eli Lilly;Merck: Consultancy. Ramachandran: Mustang Bio: Current Employment. Graf: TG Therapeutics: Research Funding; BeiGene: Research Funding; MorphoSys: Consultancy; Acerta Pharma: Research Funding. Gopal: IgM bio, BMS, merck: Research Funding; Seattle Genetics; Janssen; IMab Bio; TG Therapeutics; Astra Zeneca; Merck; Gilead; ADC Therapeutics; Nurix; TG therapeutics, Cellectar; Actinium: Consultancy; imab bio, takeda,astrazeneca,gilead: Research Funding; Seattle Genetics; Janssen; Takeda; IgM Bio; IMab Bio; BMS; Astra Zeneca; Merck; Gilead: Research Funding. Gauthier: JMP, Eusapharma, Multerra Bio: Honoraria. Cassaday: Pfizer: Honoraria, Research Funding; Seattle Genetics: Current Employment, Current equity holder in publicly-traded company; Vanda Pharmaceuticals: Research Funding; Kite/Gilead: Consultancy, Research Funding; Merck: Research Funding; Amgen: Consultancy, Research Funding. Kiem: Magenta Therapeutics, CSL,Homology Medicines, Vor Biopharma , Enochian, Umoja, Rocket Pharma: Consultancy. Turtle: Novartis: Consultancy; Myeloid Therapeutics: Current equity holder in private company, Honoraria, Membership on an entity’s Board of Directors or advisory committees; Allogene: Consultancy; PACT Pharma: Consultancy; Arsenal Bio: Current equity holder in private company, Honoraria, Membership on an entity’s Board of Directors or advisory committees; Caribou Biosciences: Current equity holder in private company, Honoraria, Membership on an entity’s Board of Directors or advisory committees; Eureka Therapeutics: Current equity holder in private company, Membership on an entity’s Board of Directors or advisory committees; Precision Biosciences: Current equity holder in publicly-traded company, Honoraria, Membership on an entity’s Board of Directors or advisory committees; Physician Education Resource: Consultancy; AstraZeneca: Consultancy, Research Funding; Nektar Therapeutics: Consultancy, Research Funding; Juno/BMS: Patents & Royalties, Research Funding; Century Therapeutics: Honoraria, Membership on an entity’s Board of Directors or advisory committees; T-CURX: Membership on an entity’s Board of Directors or advisory committees; Humanigen: Consultancy; Kite/Gilead: Consultancy. Maloney: Pharmacyclics: Consultancy, Honoraria; Juno Therapeutics: Consultancy, Honoraria, Patents & Royalties: Patents are pending, but not issued, licensed, no royalties, no licensees., Research Funding; Celgene: Consultancy, Honoraria, Research Funding; Kite, a Gilead Company: Consultancy, Honoraria, Research Funding; Gilead Sciences: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; Amgen: Consultancy, Honoraria; MorphoSys: Consultancy, Honoraria; Bioline Rx: Consultancy, Honoraria; A2 Biotherapeutics: Consultancy, Current equity holder in publicly-traded company, Honoraria; Genentech: Consultancy, Honoraria. Till: Mustang: Patents & Royalties, Research Funding

gamma-Secretase inhibitor in combination with BCMA chimeric antigen receptor T-cell immunotherapy for individuals with relapsed or refractory multiple myeloma: a phase 1, first-in-human trial

BACKGROUND: gamma-Secretase inhibitors (GSIs) increase B cell maturation antigen (BCMA) density on malignant plasma cells and enhance antitumour activity of BCMA chimeric antigen receptor (CAR) T cells in preclinical models. We aimed to evaluate the safety and identify the recommended phase 2 dose of BCMA CAR T cells in combination with crenigacestat (LY3039478) for individuals with relapsed or refractory multiple myeloma. METHODS: We conducted a phase 1, first-in-human trial combining crenigacestat with BCMA CAR T-cells at a single cancer centre in Seattle, WA, USA. We included individuals aged 21 years or older with relapsed or refractory multiple myeloma, previous autologous stem-cell transplant or persistent disease after more than four cycles of induction therapy, and Eastern Cooperative Oncology Group performance status of 0-2, regardless of previous BCMA-targeted therapy. To assess the effect of the GSI on BCMA surface density on bone marrow plasma cells, participants received GSI during a pretreatment run-in, consisting of three doses administered 48 h apart. BCMA CAR T cells were infused at doses of 50 x 10(6) CAR T cells, 150 x 10(6) CAR T cells, 300 x 10(6) CAR T cells, and 450 x 10(6) CAR T cells (total cell dose), in combination with the 25 mg crenigacestat dosed three times a week for up to nine doses. The primary endpoints were the safety and recommended phase 2 dose of BCMA CAR T cells in combination with crenigacestat, an oral GSI. This study is registered with ClinicalTrials.gov, NCT03502577, and has met accrual goals. FINDINGS: 19 participants were enrolled between June 1, 2018, and March 1, 2021, and one participant did not proceed with BCMA CAR T-cell infusion. 18 participants (eight [44%] men and ten [56%] women) with multiple myeloma received treatment between July 11, 2018, and April 14, 2021, with a median follow up of 36 months (95% CI 26 to not reached). The most common non-haematological adverse events of grade 3 or higher were hypophosphataemia in 14 (78%) participants, fatigue in 11 (61%), hypocalcaemia in nine (50%), and hypertension in seven (39%). Two deaths reported outside of the 28-day adverse event collection window were related to treatment. Participants were treated at doses up to 450 x 10(6) CAR(+) cells, and the recommended phase 2 dose was not reached. INTERPRETATIONS: Combining a GSI with BCMA CAR T cells appears to be well tolerated, and crenigacestat increases target antigen density. Deep responses were observed among heavily pretreated participants with multiple myeloma who had previously received BCMA-targeted therapy and those who were naive to previous BCMA-targeted therapy. Further study of GSIs given with BCMA-targeted therapeutics is warranted in clinical trials. FUNDING: Juno Therapeutics-a Bristol Myers Squibb company and the National Institutes of Health