Strategy to decrease off-target peptide recognition by affinity-enhanced, T cell receptor (TCR)-based T cell therapies

e14010 Background: The key driver for effective immune cell therapies is the overall binding strength of the immune cell and the target cell (e.g. tumor cells). The overall strength is known as ‘avidity’, a parameter reflecting interaction efficiency. The key to success for immune cell therapies is generating effective and long-lasting immune responses. The avidity of an immune cell to its target is predicative of its function, but current techniques to measure avidity are low-throughput and ineffective. Herein, we describe the use of acoustic forces to discriminate immune cells based on their avidity to tumor cells. The force required to separate a cell from its target is called the ‘rupture force’, and in this study, we were able to identify the rupture forces of tumor specific and non-specific T cells and enrich these different populations for downstream characterization. Methods: T cells from a healthy donor were transduced with either a non-relevant, or a melanoma recognizing T cell receptor and selected with puromycin resistance. Melanoma cells were seeded in the flow cell and allowed to adhere overnight to form a monolayer. For confocal experiments CFSE and Cell Trace far red stained T cells were mixed in a 1:1 ratio before co-culturing them in the flow cell. An acoustic force ramp was applied within the flow cell and cell detachment was monitored. Results: T cells engineered with a melanoma antigen-recognizing T-cell receptor needed 6 times more force than non-specific T cells to be separated from the melanoma target cells. Furthermore, 1.4 to 3.6-fold enrichment of high-avidity T cells was obtained from a mixed population of specific and non-specific T cells using acoustic forces. Conclusions: These findings indicate that melanoma-specific T cells bind with a higher avidity than non-specific T cells and that they can be separated with this approach. In conclusion, we demonstrate a novel method to measure cell avidity and sort cells by utilizing acoustic forces.

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Orthotopic T-Cell Receptor Replacement—An “Enabler” for TCR-Based Therapies

Cells ◽

10.3390/cells9061367 ◽

2020 ◽

Vol 9 (6) ◽

pp. 1367 ◽

Cited By ~ 4

Author(s):

Kilian Schober ◽

Thomas R. Müller ◽

Dirk H. Busch

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Technology Development ◽

Clinical Care ◽

Tumor Infiltrating Lymphocytes ◽

Cell Receptor ◽

Current Status ◽

Cell Therapies ◽

Engineered T Cells

Natural adaptive immunity co-evolved with pathogens over millions of years, and adoptive transfer of non-engineered T cells to fight infections or cancer so far exhibits an exceptionally safe and functional therapeutic profile in clinical trials. However, the personalized nature of therapies using virus-specific T cells, donor lymphocyte infusion, or tumor-infiltrating lymphocytes makes implementation in routine clinical care difficult. In principle, genetic engineering can be used to make T-cell therapies more broadly applicable, but so far it significantly alters the physiology of cells. We recently demonstrated that orthotopic T-cell receptor (TCR) replacement (OTR) by clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) can be used to generate engineered T cells with preservation of near-physiological function. In this review, we present the current status of OTR technology development and discuss its potential for TCR-based therapies. By providing the means to combine the therapeutic efficacy and safety profile of physiological T cells with the versatility of cell engineering, OTR can serve as an “enabler” for TCR-based therapies.

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Site-directed mutations in the VDJ junctional region of a T cell receptor β chain cause changes in antigenic peptide recognition

Cell ◽

10.1016/0092-8674(88)90068-2 ◽

1988 ◽

Vol 54 (4) ◽

pp. 473-484 ◽

Cited By ~ 108

Author(s):

Isaac Engel ◽

Stephen M. Hedrick

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Antigenic Peptide ◽

Peptide Recognition ◽

Junctional Region ◽

Β Chain

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Evolution of CD8+ T Cell Receptor (TCR) Engineered Therapies for the Treatment of Cancer

Cells ◽

10.3390/cells10092379 ◽

2021 ◽

Vol 10 (9) ◽

pp. 2379

Author(s):

Yimo Sun ◽

Fenge Li ◽

Heather Sonnemann ◽

Kyle R. Jackson ◽

Amjad H. Talukder ◽

...

Keyword(s):

Clinical Trials ◽

T Cell ◽

Solid Tumors ◽

T Cell Receptor ◽

Tumor Antigen ◽

Cell Receptor ◽

Patient Specific ◽

Target Antigen ◽

Tumor Escape ◽

Cell Therapies

Engineered T cell receptor T (TCR-T) cell therapy has facilitated the generation of increasingly reliable tumor antigen-specific adaptable cellular products for the treatment of human cancer. TCR-T cell therapies were initially focused on targeting shared tumor-associated peptide targets, including melanoma differentiation and cancer-testis antigens. With recent technological developments, it has become feasible to target neoantigens derived from tumor somatic mutations, which represents a highly personalized therapy, since most neoantigens are patient-specific and are rarely shared between patients. TCR-T therapies have been tested for clinical efficacy in treating solid tumors in many preclinical studies and clinical trials all over the world. However, the efficacy of TCR-T therapy for the treatment of solid tumors has been limited by a number of factors, including low TCR avidity, off-target toxicities, and target antigen loss leading to tumor escape. In this review, we discuss the process of deriving tumor antigen-specific TCRs, including the identification of appropriate tumor antigen targets, expansion of antigen-specific T cells, and TCR cloning and validation, including techniques and tools for TCR-T cell vector construction and expression. We highlight the achievements of recent clinical trials of engineered TCR-T cell therapies and discuss the current challenges and potential solutions for improving their safety and efficacy, insights that may help guide future TCR-T studies in cancer.

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On the feasibility of mining CD8+ T cell receptor patterns underlying immunogenic peptide recognition

Immunogenetics ◽

10.1007/s00251-017-1023-5 ◽

2017 ◽

Vol 70 (3) ◽

pp. 159-168 ◽

Cited By ~ 23

Author(s):

Nicolas De Neuter ◽

Wout Bittremieux ◽

Charlie Beirnaert ◽

Bart Cuypers ◽

Aida Mrzic ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cd8 T Cell ◽

Cell Receptor ◽

Peptide Recognition ◽

Immunogenic Peptide

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Implications of T cell receptor biology on the development of new T cell therapies for cancer

Immunotherapy ◽

10.2217/imt-2019-0046 ◽

2020 ◽

Vol 12 (1) ◽

pp. 89-103 ◽

Cited By ~ 1

Author(s):

Ian R Hardy ◽

Wolfgang W Schamel ◽

Patrick A Baeuerle ◽

Daniel R Getts ◽

Robert Hofmeister

Keyword(s):

T Cell ◽

Human Leukocyte Antigen ◽

T Cell Receptor ◽

Human Leukocyte ◽

Cell Receptor ◽

Tcr Signaling ◽

Cell Therapies ◽

Leukocyte Antigen ◽

Car T Cell ◽

Car T

Recently, two chimeric antigen receptor (CAR) T cell therapies were approved based on their remarkable efficacy in patients with hematological malignancies. By contrast, CAR-T cell therapies results in solid tumors have been less promising. To develop the next generation of T cell therapies a better understanding of T cell receptor (TCR) biology and its implication for the design of synthetic receptors is critical. Here, we review current and newly developed forms of T cell therapies and how their utilization of different components of the TCR signaling machinery and their requirement for engagement (or not) of human leukocyte antigen impacts their design, efficacy and applicability as cancer drugs. Notably, we highlight the development of human leukocyte antigen-independent T cell platforms that utilize the full TCR complex as having promise to overcome some of the limitations of existing T cell therapies.

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