CAR-T/Transgene/Cytokine/Biomarker RNA Biodistribution Analysis

Chimeric antigen receptor (CAR) T cells are engineered fusion proteins that target T cells to a specific antigen present on tumor cells to generate an antitumor immune response. The greater potency of CAR-T cells compared to antibody therapeutics demands a more stringent CAR-T target safety assessment to avoid adverse events resulting from "on-target/off-tumor" activity.

Furthermore, it is vital to track and monitor the pharmacokinetics of CAR+T cells within the context of intact tissue and tumor to understand the mechanisms underlying off-tumor toxicity and efficacy in tumor killing. And for IND application, the biodistribution characteristics of CAR-T cells are required to be determined using at least two methods (both quantitative and qualitative evaluation could be applicable) for a comprehensively understanding of their potential target organs/tissues.

In Situ Hybridization is an excellent method to detect the CAR-T cells, as many current methods are either not sensitive enough to detect low levels of expression or incapable of detecting CAR+ T cells in the tissue context. RNA ISH technology can be used to detect CAR, cytokines and T cell makers' expression within intact tissue morphology, addressing critical questions on tissue biodistribution, persistence, cellular tropism, and vector promoter activity. As a leading technology provider, Creative Bioarray's elite scientists have broad experience in CAR-T/Transgene/Cytokine/Biomarker RNA Biodistribution Analysis (RNA ISH method), which can supply the best service for you.

Figure 1. RNA ISH assay for detection. (A) RNA ISH probes were designed to target the 3' UTR of the CAR vector. (B) Activated CAR+ T cells traffic to the tumor site.

Features of CAR-T/Transgene/Cytokine/Biomarker RNA Biodistribution Analysis (RNA ISH method)

(1) Detection of activated CAR-T cells in the tissue context
(2) Codon-optimized transgene and biomarker expression analysis
(3) Custom probes designed within 1-2 weeks
(4) Fastest turnaround time

Benefits of CAR-T/Transgene/Cytokine/Biomarker RNA Biodistribution Analysis (RNA ISH method)

(1) Detect low levels of gene expression across all normal tissues in animal models and human samples
(2) Assess cell-specific target expression in normal tissues and expression heterogeneity in tumor tissues
(3) Visualize and quantify CAR-T cell infiltration and activation in tumor and off-tumor tissues
(4) Simultaneously detect CAR/TCR vector and cytokines, T cell markers, or other cell-type markers

Creative Bioarray offers CAR-T/Transgene/Cytokine/Biomarker RNA Biodistribution Analysis for you as follows:

• Probe design
• Probe synthesis
• ISH staining
• Imaging
• Data analysis

Quotation and ordering

Our customer service representatives are available 24hr a day! We thank you for considering Creative Bioarray as your CAR-T/Transgene/Cytokine/Biomarker RNA Biodistribution Analysis partner.

References

1. Wang F, et al. RNAscope®: A novel in situ RNA analysis platform for formalin-fixed, paraffinembedded tissues. J Mol Diagn. 2012; 14(1):22-9.
2. O'Rourke DM, et al. A single dose of peripherally infused EGFRvIII-directed CAR T cells mediates antigen loss and induces adaptive resistance in patients with recurrent glioblastoma. Sci Transl Med. 2017; 9(399): pii: eaaa0984.
3. Wing A, et al. Improving CART-cell therapy of solid tumors with oncolytic virus-driven production of a bispecific T-cell engager. Cancer Immunol Res. 2018; 6(5):605–16.