About

Bruce L. Levine, Ph.D., is the Barbara and Edward Netter Professor in Cancer Gene Therapy at the Abramson Cancer Center of the University of Pennsylvania. He is a member of the Abramson Cancer Center and serves as Deputy Director for Technology Innovation and Assessment at the Center for Cellular Immunotherapies. Levine is also the Founding Director of the Clinical Cell and Vaccine Production Facility at the University of Pennsylvania, within the Department of Pathology and Laboratory Medicine at the Perelman School of Medicine. His research expertise encompasses immunology, cell and gene therapy, with a focus on the development and manufacturing of cellular therapies. Levine's educational background includes a B.A. in Biology with a minor in History from the University of Pennsylvania and a Ph.D. in Immunology and Infectious Diseases from Johns Hopkins University. His work involves advancing cellular immunotherapies, including CAR T cell production, gene editing, and the engineering of immune cells for cancer and infectious disease treatments.

Research topics

• Immunology
• Medicine
• Internal medicine
• Biology
• Cancer research
• Genetics
• Bioinformatics
• Computational biology
• Oncology

Selected publications

• International Society for Cell & Gene Therapy Expanded Access Working Group position paper: key considerations to support equitable and ethical expanded access to investigational cell- and gene-based interventions

  Cytotherapy · 2025-02-07 · 4 citations

  articleOpen access

  This position paper reviews the Expanded Access pathway for cell and gene therapies, examining its critical role at the nexus of patient need, regulatory frameworks, and scientific advancement. Spearheaded by the International Society for Cell & Gene Therapy's Expanded Access Working Group, it explores how investigational therapies are accessed outside of clinical trials for patients with serious or life-threatening conditions when no approved alternatives exist. Access to cell and gene therapy products are of specific interest to patients because many times the products are bespoke, being used to treat serious and/or incurable conditions, and are potentially curative. As the field of cell and gene therapy rapidly progresses, healthcare professionals face mounting challenges in navigating the balance between access and oversight. Key considerations include transparent communication with patients, robust data reporting, and a discussion of cost recovery models and their implications for long-term commercialization strategies. Equity and inclusivity are central themes, highlighting the need to design pathways that are accessible to diverse patient populations while upholding high scientific and ethical standards. This position paper is presented as a resource for clinicians, researchers, and policymakers navigating the evolving landscape of investigational cell and gene therapies. It emphasizes the importance of ethical frameworks and equitable practices in delivering transformative treatments to patients in need.

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• Long-term safety of lentiviral or gammaretroviral gene-modified T cell therapies

  Nature Medicine · 2025-01-20 · 69 citations

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• Engineering Relapse-Resistant CAR T-Cells for Blood Cancers (Penta T-Cells)

  Cytotherapy · 2025-04-30

  articleOpen access

  Immunotherapy with chimeric antigen receptor (CAR) T-cells has demonstrated remarkable success in treating hematological malignancies . Despite impressive outcomes, relapse driven by antigen-negative disease remains a critical challenge. To overcome tumor antigen escape, effector T-cells are engineered to co-express two CARs with distinct antigen specificities such as CD19+CD20 or CD19+CD22. However, tumor relapses still occur and limitations in the manufacturing of multi-receptor CAR T-cells restrict clinical translation, highlighting the need for innovative approaches. Human T cells were expanded from healthy donor PBMCs and transduced with lentiviral vectors encoding CTL019 (anti-CD19 CAR) or Penta (anti-CD19/20/22/79b CAR and minimal low-affinity nerve growth factor receptor , mLNGFR). Surface marker expression was assessed via flow cytometry. Cytotoxic assays were performed with Burkitt lymphoma cells (Ramos) expressing a luciferase reporter. Antigen knockout models were generated via ribonucleoprotein (RNP)-mediated CRISPR-Cas9 genome editing. We developed a platform for engineering and manufacturing multi-receptor CAR T-cells accommodating transgenes beyond current size limitations. As proof of concept, we generated the first T cell with five independent receptors based on a single-vector design (Penta T-cell): four CARs to enable simultaneous targeting of CD19/CD20/CD22/CD79b and mLNGFR for monitoring (Fig. 1A). In vitro studies using single-antigen-positive Ramos tumor models validate that, in contrast to monospecific anti-CD19 CAR T-cells, the cytotoxic activity of Penta T-cells is not limited to CD19+ cancers but extends to CD20+, CD22+, and CD79b+ tumors (Fig. 1B). Moreover, by combining single-antigen-positive tumors to create an antigen-heterogeneous cancer model (mixed CD19+/CD20+/CD22+/CD79b+ tumor), we demonstrate that Penta T-cells effectively clear heterogeneous cancers by targeting all four antigens simultaneously (Figs. 1B and 1C). In contrast, anti-CD19 CAR T-cells eliminate only the CD19+ tumor fraction, resulting in antigen-escape. We successfully engineered effector T cells with five independent receptors and demonstrated their enhanced cytotoxicity against heterogeneous tumors in vitro. Safety and efficacy studies with human tumor xenograft models are ongoing. Our work represents a crucial proof-of-concept for relapse-resistant CAR T-cells, addressing current barriers in CAR T-cell design and production.

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• Assessing the oncogenic risk: the long-term safety of autologous chimeric antigen receptor T cells

  The Lancet · 2025-02-27 · 11 citations

  review
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• CCR5-targeted allogeneic gamma–delta CD19 chimeric antigen receptor T cells for HIV-associated B cell-malignancy immunotherapy

  Nature Biomedical Engineering · 2025-10-21 · 1 citations

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• Improving American chestnut resistance to two invasive pathogens through genome-enabled breeding

  bioRxiv (Cold Spring Harbor Laboratory) · 2025-02-01 · 2 citations

  preprintOpen access

  Over a century after two introduced pathogens decimated American chestnut populations, breeding programs continue to incorporate resistance from Chinese chestnut to recover self-sustaining populations. Due to complex genetics of chestnut blight resistance, it is challenging to obtain trees with sufficient resistance and competitive growth. We developed high quality reference genomes for Chinese and American chestnut and leveraged large disease phenotype and genotype datasets to develop accurate genomic selection. Inoculation and simulation results indicate that resistance may be substantially increased in trees that inherited 70% to 100% of their genome from American chestnut. To facilitate gene editing, we integrated multiple lines of evidence to discover candidate alleles for blight resistance and susceptibility. These genomic resources provide a strong foundation to accelerate restoration of this iconic tree.

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• Multiplex engineering using microRNA-mediated gene silencing in CAR T cells

  Frontiers in Immunology · 2025-08-21 · 4 citations

  articleOpen accessSenior authorCorresponding

  Background Multiplex gene-edited chimeric antigen receptor (CAR) T-cell therapies face significant challenges, including potential oncogenic risks associated with double-strand DNA breaks. Targeted microRNAs (miRNAs) may provide a safer, functional, and tunable alternative for gene silencing without the need for DNA editing. Methods As a proof of concept for multiplex gene silencing, we employed an optimized miRNA backbone and gene architecture to silence T-cell receptor (TCR) and major histocompatibility complex class I (MHC-I) in mesothelin-directed CAR (M5CAR) T cells. The efficacy of this approach was compared to CD3ζ and β2-microglobulin (β2M) CRISPR/Cas9 knockout (KO) cells. miRNA-expressing cassettes were incorporated into M5CAR lentiviral vectors, enabling combined gene silencing and CAR expression. Antitumor activity was evaluated using in vitro assays and in vivo pancreatic ductal adenocarcinoma models. Results Silenced (S) M5CAR T cells retained antitumor functionality comparable to, and in some cases exceeding, that of KO cells. In vivo , S M5CAR T cells achieved tumor control with higher persistence and superior metastasis prevention. In vitro assays demonstrated enhanced resistance to alloreactive natural killer (NK) cells and peripheral blood mononuclear cells (PBMCs). Conclusions Titratable multiplex gene silencing via targeted miRNAs offers an alternative to gene editing for CAR T cells, with potential advantages in potency, persistence, metastasis prevention, and immune evasion for allogeneic products. This strategy may overcome tumor-induced immunosuppression while avoiding the risks associated with DNA double-strand breaks.

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• Cellular gene therapy access in Africa: a multifactorial feasibility analysis for implementation

  Cytotherapy · 2025-04-17 · 3 citations

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• Humanized CD19 chimeric antigen receptor (CAR) T-cell therapy for high-risk and post-CAR relapse of B-cell acute lymphoblastic leukemia

  Blood · 2025-11-03

  article

  Abstract Background CD19-directed chimeric antigen receptor T-cell (CART19) therapy has transformed the treatment landscape for relapsed/refractory (r/r) B-cell acute lymphoblastic leukemia (B-ALL). Yet relapse occurs in approximately 50% of recipients. These patients and subgroups of pediatric patients in first relapse, currently ineligible for commercial CART19, have very poor outcomes with current approaches, warranting investigation of alternative strategies. We previously investigated a humanized CART19 (huCART19) in a Phase I trial with encouraging responses in both CAR-naïve and CAR-exposed cohorts (Myers JCO 2021). Here we report outcomes from a Phase II clinical trial (NCT03792633) of huCART19 for high risk r/r B-ALL, including early bone marrow (BM) relapse, a subgroup with historically poor outcomes despite intensive therapy. Methods Patients aged 0-29 years (y) with CD19+ B-ALL were eligible in 2 cohorts: CAR-naïve patients with B-ALL that is refractory, in high risk first relapse, second or greater relapse, or relapsed after or ineligible for hematopoietic stem cell transplant (HSCT); CAR-exposed patients with poor response to prior cell therapy. Patients received huCART19 at a dose of 5x106 CAR T cells/kg (maximum 2.5x108) after lymphodepletion (LD) with fludarabine and cyclophosphamide. Response was assessed on day 28 by BM and cerebrospinal fluid morphology, minimal residual disease (MRD) by flow cytometry, and biologic response by B cell aplasia (BCA). The primary endpoint was event-free survival (EFS). Secondary endpoints included overall response rate (ORR), defined as rate of complete remission (CR) or CR with incomplete hematologic recovery (CRi) with BCA, and relapse-free survival (RFS). Data cutoff was July 1, 2025. Results Of 106 patients enrolled, 100 were infused with huCART19 from 3/2019-8/2023. The median age at infusion was 12y (range 1-29), 44% were female, 20% Hispanic, 5% Asian, 7% Black, and 6% had Trisomy 21. Prior therapy included HSCT in 21%, blinatumomab in 21% and inotuzumab in 15%. The CAR-naïve cohort (n=52) included 25 with first early BM relapse within 36m of diagnosis (12 <18m). On pre-infusion BM performed post-LD, 13/52 (25%) had >25% blasts, 25/52 (48%) <0.01%. Three patients were inevaluable for response: 1 deemed ineligible due to myeloid lineage switch on pre-infusion BM; 1 died on day 2; 1 lost to follow-up at day 28. By day 28, 46/49 (94%) were in CR/CRi, 45/46 MRD-negative, 1 MRD inevaluable. With a median follow up of 51m, EFS and RFS at 2y were 65% (95% CI 52-81%) and 70% (95% CI 57-86%), and at 4y, 57% (95% CI 42-76%) and 62% (95% CI 47-81%). Relapse occurred in 14/46 (30%), of which 7 (50%) were CD19-. Ten patients pursued alternative therapy in remission, 7 for loss of BCA and 3 for MRD recurrence (2 CD19+, 1 CD19-). Overall survival (OS) at 2y and 4y was 74% (95% CI 63-87%). In a subgroup analysis of CAR-naïve patients treated for early BM relapse, ORR was 21/23 (91%), 2y EFS was 39% (95% CI 21-72%), RFS 45% (95% CI 25-81%), and OS 57% (95% CI 40-81%). The CAR-exposed cohort (n=48) included 20 with post-CART19 relapse and 28 with early (<6m) loss of BCA without relapse. On pre-infusion BM, 5/48 (10%) had >25% blasts, 34/48 (71%) <0.01%. By day 28, 43/48 patients were in CR/CRi, 43/43 MRD-negative; 6/43 did not establish BCA resulting in an ORR of 37/48 (77%). With a median follow up of 43m, EFS and RFS at 2y were 53% (95% CI 39-71%) and 72% (95% CI 57-91%), and at 4y, 50% (95% CI 36-68%) and 68% (95% CI 52-88%). Relapse occurred in 9/37 (24%), of which 4 (44%) were CD19-. Eleven patients received alternative therapy in remission, 6 for loss of BCA, 4 for CD19+ MRD recurrence, and 1 for therapy-related myeloid neoplasm. OS at 2y and 4y was 77% (95% CI 66-90%). CRS was reported in 47/52 (90%, 10 grade [Gr] 3, 5 Gr 4 on Penn scale) CAR-naïve patients and 38/48 (79%, 1 Gr 3, 2 Gr 4) CAR-exposed. There was 1 death prior to day 28, due to gastrointestinal hemorrhage on day 2 in the setting of progressive ALL and Gr 2 CRS. CAR neurotoxicity was reported in 14/52 (27%, 2 Gr 3, 2 Gr 4) CAR-naïve patients and 7/48 (15%, 1 Gr 3, 1 Gr 4) CAR-exposed, with 1 case of Gr 3 cerebral edema, fully recovered, and 1 case of ongoing myelopathy. Conclusions HuCART19 produced durable remissions in high risk r/r B-ALL and demonstrated efficacy as salvage therapy for those with poor response to prior CAR therapy, comparing favorably to historical outcomes in this extremely high risk group.

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• T cell lymphoma and secondary primary malignancy risk after commercial CAR T cell therapy

  Nature Medicine · 2024 · 250 citations

  • Medicine
  • Oncology
  • Internal medicine
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Recent grants

• NIH Grant R01CA166961

  NIH · $960k · 2017

• Enhancing Chimeric Antigen Receptor T Cell Therapies for HematologicMalignancies: Beyond CART 19

  NIH · $39.1M · 2017–2028

Frequent coauthors

• Carl H. June

  Parker Institute for Cancer Immunotherapy

  927 shared

• Simon F. Lacey

  University of Pennsylvania

  175 shared

• Stephan A. Grupp

  Children's Hospital of Philadelphia

  168 shared

• J. Joseph Melenhorst

  Cleveland Clinic

  167 shared

• Zhaohui Zheng

  Xijing Hospital

  140 shared

• Bruce R. Blazar

  University of Minnesota

  133 shared

• Julio Cotte

  University of Pennsylvania

  121 shared

• Stephen J. Schuster

  University of Pennsylvania

  119 shared