A5025983003- CAR-T cell therapy research
- Virus-based gene therapy research
- Immunotherapy and Immune Responses
- Immune Cell Function and Interaction
- CRISPR and Genetic Engineering
- Viral Infectious Diseases and Gene Expression in Insects
- T-cell and B-cell Immunology
- RNA Interference and Gene Delivery
- Nanowire Synthesis and Applications
- Silicon Carbide Semiconductor Technologies
- Cancer Immunotherapy and Biomarkers
- Cancer, Hypoxia, and Metabolism
- Monoclonal and Polyclonal Antibodies Research
- HIV Research and Treatment
- Biosimilars and Bioanalytical Methods
- Estrogen and related hormone effects
- Cytomegalovirus and herpesvirus research
- Angiogenesis and VEGF in Cancer
- Chronic Lymphocytic Leukemia Research
- Cancer, Lipids, and Metabolism
- Cancer-related molecular mechanisms research
- Alzheimer's disease research and treatments
- NF-κB Signaling Pathways
- BRCA gene mutations in cancer
- Chemokine receptors and signaling
University of Pennsylvania
2014-2023
Takeda (United States)
2023
Abramson Cancer Center
2020
Cancer Research Institute
2010-2015
Cancer Research Center
2015
UPMC Hillman Cancer Center
2010-2014
Sichuan University
2002-2014
West China Hospital of Sichuan University
2002-2014
Duke Medical Center
2003-2011
National Institutes of Health
2005-2010
CRISPR-Cas9 gene editing provides a powerful tool to enhance the natural ability of human T cells fight cancer. We report first-in-human phase 1 clinical trial test safety and feasibility multiplex engineer in three patients with refractory Two genes encoding endogenous cell receptor (TCR) chains, TCRα (
Abstract Purpose: Using gene-disrupted allogeneic T cells as universal effector provides an alternative and potentially improves current chimeric antigen receptor (CAR) T-cell therapy against cancers infectious diseases. Experimental Design: The CRISPR/Cas9 system has recently emerged a simple efficient way for multiplex genome engineering. By combining lentiviral delivery of CAR electro-transfer Cas9 mRNA gRNAs targeting endogenous TCR, β-2 microglobulin (B2M) PD1 simultaneously, to...
Off-target toxicity due to the expression of target antigens in normal tissue represents a major obstacle use chimeric antigen receptor (CAR)-engineered T cells for treatment solid malignancies. To circumvent this issue, we established clinical platform engineering with transient CAR by using vitro transcribed mRNA encoding that includes both CD3-ζ and 4-1BB co-stimulatory domains. We present two case reports from ongoing trials indicating adoptive transfer mesothelin (CARTmeso cells) is...
T cells can be redirected to overcome tolerance cancer by engineering with integrating vectors express a chimeric antigen receptor (CAR). In preclinical models, we have previously demonstrated that transfection of messenger RNA (mRNA) coding for CAR is an alternative strategy has antitumor efficacy and the potential evaluate on-target off-tumor toxicity new targets safely due transient mRNA expression. Here, report safety observed in four patients treated autologous had been electroporated...
Target-mediated toxicity is a major limitation in the development of chimeric antigen T-cell receptors (CAR) for adoptive cell therapy solid tumors. In this study, we developed strategy to adjust affinities scFv component CAR discriminate tumors overexpressing target from normal tissues that express it at physiologic levels. A CAR-expressing panel was generated with varying over three orders magnitude. High-affinity cells recognized expressed any level, including levels were undetectable by...
Cancer has an impressive ability to evolve multiple processes evade therapies. While immunotherapies and vaccines have shown great promise, particularly in certain solid tumors such as prostate cancer, they been met with resistance from that use a multitude of mechanisms immunosuppression limit effectiveness. Prostate particular, secretes transforming growth factor β (TGF-β) means inhibit immunity while allowing for cancer progression. Blocking TGF-β signaling T cells increases their...
Abstract Chimeric antigen receptor (CAR)–modified adoptive T-cell therapy has been successfully applied to the treatment of hematologic malignancies, but faces many challenges in solid tumors. One major obstacle is immune-suppressive effects induced both naturally occurring and genetically modified tumor-infiltrating lymphocytes (TIL) by inhibitory receptors (IR), namely PD1. We hypothesized that interfering with PD1 signaling would augment CAR activity against To address this possibility,...
Abstract Little is known about the biology of murine T-cell receptors (TCR) expressed in human cells. We recently observed that a anti-human p53 TCR highly functional when lymphocytes. Herein, we compare and mouse function expression to delineate molecular basis for apparent superior biological activity T To this end, created hybrid TCRs where swapped original constant regions with either or ones, respectively. showed “murinized” were overexpressed on surface lymphocytes compared their...
Redirecting T lymphocyte antigen specificity by gene transfer can provide large numbers of tumor-reactive lymphocytes for adoptive immunotherapy. However, safety concerns associated with viral vector production have limited clinical application cells expressing chimeric receptors (CAR). be modified RNA electroporation without integration-associated concerns. To establish a safe platform immunotherapy, we first optimized the backbone in vitro transcription to achieve high-level transgene...
The effects of transgenically encoded human and mouse IL-18 on T cell proliferation its application in boosting chimeric antigen receptor (CAR) cells are presented. Robust enhancement IL-18-secreting occurred a xenograft model, this was dependent TCR IL-18R signaling. augmented IFN-γ secretion activated by the endogenous TCR. TCR-deficient, IL-18-expressing CD19 CAR exhibited enhanced antitumor activity model. Antigen-propelled activation cytokine helper ensemble (APACHE) displayed inducible...
Chimeric antigen receptors (CAR) are synthetic molecules that provide new specificities to T cells. Although successful in treatment of hematologic malignancies, CAR cells ineffective for solid tumors date. We found the cell-surface molecule c-Met was expressed ∼50% breast tumors, prompting construction a cell specific c-Met, which halted tumor growth immune-incompetent mice with xenografts. then evaluated safety and feasibility treating metastatic cancer intratumoral administration...
T cells can be engineered to express the genes of chimeric antigen receptors (CARs) that recognize tumor-associated antigens. We constructed and compared 2 CARs contained a single chain variable region moiety recognized CD19. One CAR signaling 4-1BB molecule other did not. selected not contain for further preclinical development. demonstrated gammaretroviruses encoding this receptor could transduce human cells. Anti-CD19-CAR-transduced CD8+ CD4+ produced interferon-γ interleukin-2...
// Jiangtao Ren 1 , Xuhua Zhang Xiaojun Liu Chongyun Fang Shuguang Jiang Carl H. June 1, 2 Yangbing Zhao Center for Cellular Immunotherapies, Perelman School of Medicine, University Pennsylvania, Philadelphia, PA, USA Department Pathology and Laboratory Correspondence to: Zhao, email: yangbing@upenn.edu Keywords: CRISPR/CAS9, T lymphocytes, chimeric antigen receptors, PD-1, CD95 Received: November 18, 2016 Accepted: January 27, 2017 ...
Abstract Single and dual amino acid substitution variants were generated in the TCR CDRs of three TCRs that recognize tumor-associated Ags. Substitutions enhance reactivity gene-modified T cells to cognate Ag complex identified using a rapid RNA-based transfection system. The screening panel 1G4 TCR, recognizes peptide corresponding residues 157–165 human cancer testis NY-ESO-1 (SLLMWITQC) context HLA-A*02 class I allele, resulted identification single CDR3α CDR2β substitutions dramatically...
Abstract To generate chimeric Ag receptors (CARs) for the adoptive immunotherapy of cancer patients with ErbB2-expressing tumors, a single-chain Ab derived from humanized mAb 4D5 Herceptin (trastuzumab) was initially linked to T cell signaling domains CD28 and CD3ζ CAR against ErbB2. Human PBLs expressing demonstrated Ag-specific activities ErbB2+ tumors. However, gradual loss transgene expression noted transduced this CAR. When domain truncated or mutated, not observed, suggesting that...
This study compared second-generation chimeric antigen receptors (CAR) encoding signaling domains composed of CD28, ICOS, and 4-1BB (TNFRSF9). Here, we report that certain CARs endow T cells with the ability to undergo long-term autonomous proliferation. Transduction primary human lentiviral vectors some resulted in sustained proliferation for up 3 months following a single stimulation through T-cell receptor (TCR). Sustained numeric expansion was independent cognate did not require addition...
The use of nonviral gene transfer methods in primary lymphocytes has been hampered by low efficiency and high transfection-related toxicity. In this report, transfection with toxicity was achieved electroporation using vitro-transcribed mRNA. Using these methods, >90% transgene expression >80% viable cells observed stimulated human murine T transfected GFP or mCD62L. Electroporation unstimulated PBMCs splenocytes RNA yielded 95 56% GFP+ cells, respectively. mRNA for NY-ESO-1, MART-1, p53...
Immunotherapies based on adoptive cell transfer are highly effective in the treatment of metastatic melanoma, but use this approach other cancer histologies has been hampered by identification appropriate target molecules. Immunologic approaches targeting tumor vasculature provide a means for therapy multiple solid types. We developed method to vasculature, using genetically redirected syngeneic or autologous T cells. Mouse and human cells were engineered express chimeric antigen receptor...
Cytotoxic T lymphocytes (CTLs) modified with chimeric antigen receptors (CARs) for adoptive immunotherapy of hematologic malignancies are effective in preclinical models and being tested several clinical trials. Although CTLs bearing stably expressed CARs generated by integrating viral vectors efficacious have potential long-term persistence, this mechanism CAR expression can potentially result significant toxicity. cells were electroporated an optimized vitro transcribed RNA encoding a...
Abstract We examined the activity of human T cells engineered to express variants a single TCR (1G4) specific for cancer/testis Ag NY-ESO-1, generated by bacteriophage display with wide range affinities (from 4 μM 26 pM). CD8+ expressing intermediate- and high-affinity 1G4 bound NY-ESO-1/HLA-A2 tetramers high avidity specificity, but increased affinity was associated loss target cell specificity gene-modified cells. highest (KD value pM) completely lost specificity. The TCRs in midrange, KD...
Abstract cDNAs encoding TCR α- and β-chains specific for HLA-A2-restricted cancer-testis Ag NY-ESO-1 were cloned using a 5′RACE method from RNA isolated CTL generated by in vitro stimulation of PBMC with modified NY-ESO-1-specific peptide (p157–165, 9V). Functionality the was confirmed electroporation primary PBL. cDNA these used to construct murine stem cell virus-based retroviral vector, high titer packaging lines generated. Gene transfer efficiency T lymphocytes up 60% obtained without...