expansion-enrichment-gamma-delta-t-cells-from-apheresed-human

Expansion and Enrichment of Gamma-Delta (&#947;&#948;) T Cells from Apheresed Human Product

Although V&#947;9V&#948;2 T cells are a minor subset of T lymphocytes, this population is sought after for its ability to recognize antigens in a major ...

Moffitt Cancer Center and Research Institute

Chimeric antigen receptor (CAR)-modified T cells have generated broad interest in oncology following a series of dramatic clinical successes in patients with chemorefractory B cell malignancies. CAR therapy now appears to be on the cusp of regulatory approval as a cell-based immunotherapy. We review here the T cell biology and cell engineering research that led to the development of second generation CARs, the selection of CD19 as a CAR target, and the preclinical studies in animal models that laid the foundation for clinical trials targeting CD19+ malignancies. We further summarize the status of CD19 CAR clinical therapy for non-Hodgkin lymphoma and B cell acute lymphoblastic leukemia, including their efficacy, toxicities (cytokine release syndrome, neurotoxicity and B cell aplasia) and current management in humans. We conclude with an overview of recent pre-clinical advances in CAR design that argues favorably for the advancement of CAR therapy to tackle other hematological malignancies as well as solid tumors. 

Biology and clinical application of CAR T cells for B cell malignancies.

T cells kill microbial-infected and malignant cells by detection of nonself antigens with the TCR. Tumor reactivity can be encoded genetically by introducing a chimeric antigen receptor (CAR) into T cells. CARs are composed of an antigen-binding domain and an intracellular T cell activation domain. Early human trials evaluating CD19-targeted CAR T cells for chronic lymphocytic leukemia (CLL) showed limited responses until CARs included a costimulation domain, and conditioning chemotherapy was given before T cell infusion. Clinical trials evaluating CD19-targeted CAR T cells for B cell acute lymphoblastic leukemia (B-ALL) are demonstrating response rates up to 90%. However, these clinical outcomes are associated with a cytokine release syndrome (CRS), which is caused by T cell activation and manifests as high-grade fever, hypotension, and other cardiovascular complications. It is currently managed conservatively but can be treated with cytokine-directed therapy or with high-dose steroids. Current efforts are dedicated to confirming the clinical efficacy and managing toxicities in multicenter Phase II trials. We present a thorough overview of the preclinical and clinical development of CAR T cell therapy that will highlight important areas for the basic researcher to investigate in the laboratory and contribute to this exciting field.

At The Bedside: Clinical review of chimeric antigen receptor (CAR) T cell therapy for B cell malignancies.

Adoptive T cell therapy has demonstrated tremendous outcomes against treatment-refractory leukemias and solid tumor malignancies. As opposed to industry-developed drugs that are manufactured and dispensed to hospitals and/or patients, T cells are produced in academic laboratories for clinical research and are a highly personalized therapy that represents a "living drug." The technology behind genetic modification of primary T cells has been developed and refined by a few academic medical centers. We anticipate that the exciting results generated by these efforts will lead to further investigation by other academic and industry institutions. To facilitate this adaptation we present optimized protocols for gammaretroviral production, T cell isolation, and genetic modification to create gene-targeted T cells.

Gammaretroviral Production and T Cell Transduction to Genetically Retarget Primary T Cells Against Cancer.

Myelodysplastic syndromes (MDSs) and related myeloid neoplasms represent heterogeneous diseases with overall poor outcomes related to lack of efficacious agents. Immunotherapy has revolutionized the treatment paradigm in solid malignancies and select hematologic malignancies with durable remissions in treatment-refractory populations. The initial clinical studies using immunotherapy in MDS are under way, although significant efforts are required in which to better understand their role in this disease. Likely, combinations of therapies will be required to improve outcomes in this patient population. Furthermore, identification of novel targets for antibody or T-cell modulation strategies is a matter of intensive investigation and ideally will provide treatment options in the near future for our patients.

Is Disease-Specific Immunotherapy a Potential Reality for MDS?

NKG2D-based chimeric antigen receptor therapy induced remission in a relapsed/refractory acute myeloid leukemia patient.

CAR T cells, immunologic and cellular therapies in hematologic malignancies.

One of the challenges of adoptive T-cell therapy is the development of immune-mediated toxicities including cytokine release syndrome (CRS) and neurotoxicity (NT). We aimed to identify factors that place patients at high risk of severe toxicity or treatment-related death in a cohort of 75 patients with large B-cell lymphoma treated with a standard of care CD19 targeted CAR T-cell product (axicabtagene ciloleucel).

Tumor Microenvironment Composition and Severe Cytokine Release Syndrome (CRS) Influence Toxicity in Patients with Large B-Cell Lymphoma Treated with Axicabtagene Ciloleucel.

An obstacle to the development of chimeric antigen receptor (CAR) T cells is the limited understanding of CAR T-cell biology and the mechanisms behind their antitumor activity. We and others have shown that CARs with a CD28 costimulatory domain drive high T-cell activation, which leads to exhaustion and shortened persistence. This work led us to hypothesize that by incorporating null mutations of CD28 subdomains (YMNM, PRRP, or PYAP), we could optimize CAR T-cell costimulation and enhance function. , we found that mice given CAR T cells with only a PYAP CD28 endodomain had a significant survival advantage, with 100% of mice alive after 62 days compared with 50% for mice with an unmutated endodomain. We observed that mutant CAR T cells remained more sensitive to antigen after  antigen and PD-L1 stimulation, as demonstrated by increased cytokine production. The mutant CAR T cells also had a reduction of exhaustion-related transcription factors and genes such as , and  Our results demonstrated that CAR T cells with a mutant CD28 endodomain have better survival and function. This work allows for the development of enhanced CAR T-cell therapies by optimizing CAR T-cell costimulation.

Article	Year
Biology and clinical application of CAR T cells for B cell malignancies. International journal of hematology\| PubMed ID: 27262700	2016
At The Bedside: Clinical review of chimeric antigen receptor (CAR) T cell therapy for B cell malignancies. Journal of leukocyte biology\| PubMed ID: 27354412	2016
Gammaretroviral Production and T Cell Transduction to Genetically Retarget Primary T Cells Against Cancer. Methods in molecular biology (Clifton, N.J.)\| PubMed ID: 27787796	2017
Is Disease-Specific Immunotherapy a Potential Reality for MDS? Clinical lymphoma, myeloma & leukemia\| PubMed ID: 28760299	2017
NKG2D-based chimeric antigen receptor therapy induced remission in a relapsed/refractory acute myeloid leukemia patient. Haematologica\| PubMed ID: 29703727	2018
CAR T cells, immunologic and cellular therapies in hematologic malignancies. Best practice & research. Clinical haematology\| PubMed ID: 29909911	2018
Tumor Microenvironment Composition and Severe Cytokine Release Syndrome (CRS) Influence Toxicity in Patients with Large B-Cell Lymphoma Treated with Axicabtagene Ciloleucel. Clinical cancer research : an official journal of the American Association for Cancer Research\| PubMed ID: 32669372	2020
CD28 Costimulatory Domain-Targeted Mutations Enhance Chimeric Antigen Receptor T-cell Function. Cancer immunology research\| PubMed ID: 33188139	2021

Marco Davila

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