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Viability and Bio-distribution Study of CAR-T

To date, CAR-T cell immunotherapy has received tremendous success for treating hematological malignancies (e.g., anti-CD19 CAR-T cells in leukemias), while not yet extrapolated to solid tumors. A major obstacle associated with current CAR-T cell therapy is that the limited replicative lifespan of CAR-T cells prohibits the in vivo expansion and long-term persistence of these cells, potentially hindering the long-term therapeutic efficacy of CAR-T cells. Many approaches have been evaluated such as adding signaling moieties from co-stimulatory molecules (the 2nd and 3rd generation of CAR), using T cell recognizing Epstein-Bar virus antigens and selecting central memory T cells for genetic modification.

However, CAR-T cell therapy may cause some acute on-target/on-tumor toxicity (cytokine release syndrome, tumor lysis syndrome) or on-target/off-tumor toxicity, and in some cases, CAR-T cells undergo antigen-independent proliferation that could increase cell-mediated toxicity and raise the concern of immortalization of infused CAR-T cells. Therefore, the 4th generation of CAR has been developed to address these problems, for example, harboring an inducible suicide gene in the CAR construct.

As a frontier biotech service provider, Creative Biolabs now provides preclinical viability and bio-distribution analysis services for CAR-T cell therapy. All tests are conducted by experienced technicians on the most appropriate models with the most advanced techniques.

Our studies focus on but not limited to the following aspects.

In Vivo Proliferation, Survival, and Long-Term Persistence of CAR-T Cells

  • Elucidating the proper delivery route, dosing regimen and durability of effects

T Cell Migration and Distribution

  • Ultimately determine the cell fate
  • Migration to tumor tissue is essential for anti-tumor effect
  • Migration to non-tumor site may be associated with adverse effect

Memory T Cell Response

  • T cell phenotype analysis
  • T cell response to re-challenge with tumor cells

T Cell Perseverance
The tumor microenvironment contains multiple inhibitory factors to potentially suppress CAR-T cells. Tumor cells also develop many mechanisms to avoid the eradication by CAR-T cells.

  • CD4+CD25hiFoxP3+ regulatory T cells (Tregs) analysis
  • Myeliod-derived suppressor cell (MDSCs) analysis
  • Immune checkpoint (PD-L1) analysis
  • T cell suppressive cytokine (TGF-β and IL-10) analysis

Efficacy of Next Generation CAR

  • Multiple assays on viability and bio-distribution characteristics of your novel CAR constructs depending on your purpose

Techniques Involved

  • Bioluminescence imaging of CAR-T cells
  • Flow cytometry analysis of CAR-T cells
  • Quantitative PCR
  • Histologic analysis

Immunosuppressive Tumor Microenvironment

Immunosuppressive Tumor Microenvironment
Newick K et al. Mol Ther Oncolytics, 2016

All studies are performed in a GLP-compliant and IACUC-regulated facility. We are glad to share our valuable experience on the study of viability and bio-distribution of CAR-T cell with our clients. Scientists of Creative Biolabs will work with you to design the program that best fits your requirements and expedite your CAR-T therapy to clinical trials.

All services and products are only for lab research use, not for any clinical diagnosis or treatment.

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