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

RNA In Situ Hybridization (RNA ISH) assay from Creative Bioarray is a valuable technique in CAR-NK/transgene/Cytokine/Biomarker RNA Biodistribution analysis, especially in the context of cell therapy development.

The use of RNA In Situ Hybridization (RNA ISH) technique in CAR-NK/transgene/Cytokine/Biomarker RNA Biodistribution Analysis offers several advantages:

• Cellular Resolution: RNA ISH assay allows for the visualization of RNA expression within individual cells. This is particularly advantageous when determining viral biodistribution as it allows for the identification of the types of cells that are being infected or targeted.
• Quantitative Analysis: With RNA ISH assay, quantification of RNA molecules is possible. This feature is significant in biodistribution analysis as it helps in understanding the level of gene expression among different cells accurately.
• Spatial Information: RNA ISH assay helps to locate exactly where the molecule is expressed in the tissue, thereby providing valuable spatial information for understanding the biological function of each cell type.
• Highly Specific: The technique is highly specific, allowing for the detection of specific RNA sequences among many. For instance, it can be used to detect CAR-NK cell biodistribution in the tissues.
• Simultaneous Detection: RNA ISH assay allows for simultaneous detection of multiple targets, thus enabling the detection of various biomarkers, cytokines, and transgene at the same time.
• Tissue Preservation: Unlike other approaches like PCR, with RNA ISH assay tissue morphology is well preserved which is crucial in biodistribution analysis.

While traditionally RNA ISH has been associated with limitations such as sensitivity, the development of novel techniques such as RNAscope ISH has greatly improved this tool, providing an exceptional approach to CAR-NK/transgene/Cytokine/Biomarker RNA Biodistribution Analysis.

Fig. 1. RNA ISH assay for detection. RNA ISH probes were designed to target the 3' UTR of the CAR vector.

Fig. 2 RNAscope assay of CAR-T and TCR-T cells. (Left) ROR1-targeted CAR-T cells infiltration and activation in ROR1-positive mouse lung and liver. RNAscope CAR 3’UTR probe and GZMB probe detect activated CAR+ T cell; Mm-Ror1 probe signal in mouse liver and lung. (Middle) BCMA-targeted CAR-T cell infiltration and activation in BCMA-positive xenograft tumor. RNAscope CAR 3'UTR and IFNG probe with CD3 immunofluorescence.

The importance of biodistribution testing for CAR-NK technology lies in the evaluation of the biological behavior and safety levels of these therapeutic cells once they are introduced into the body. Through this, researchers can assess where these cells travel, how long they persist, which organs they target, and also monitor any potential toxicity or side effect profiles.

Biodistribution can influence the efficacy of the therapy and if it's not properly assessed, it could lead to off-target effects which may impact the safety of patients. Thus, it is an essential part of the preclinical evaluation of cell therapies like CAR-NK to ensure controlled and directed outcomes when they are used in human subjects.

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

• Detection of activated CAR-NK cells in the tissue context
• Codon-optimized transgene and biomarker expression analysis
• Custom probes designed within 1-2 weeks
• Fastest turnaround time

Benefits of CAR-NK/transgene/Cytokine/Biomarker RNA Biodistribution Analysis (RNA ISH method)

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

Creative Bioarray offers CAR-NK/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-NK/Transgene/Cytokine/Biomarker RNA Biodistribution Analysis partner.

References

1. Wright JH, Huang L-Y, Weaver S et al. (2021) Detection of engineered T cells in FFPE tissue by multiplex in situ hybridization and immunohistochemistry. Journal of Immunological Methods 492:112955.
2. Gauthier, L., et al. (2019). Multifunctional Natural Killer Cell Engagers Targeting NKp46 Trigger Protective Tumor Immunity. Cell. 177(7):1701-1713
3. Ramachandran I., et al. (2019). Systemic and local immunity following adoptive transfer of NY-ESO-1 SPEAR T cells in synovial sarcoma. J Immunother Cancer 7:276
4. Tang N, Cheng C, Zhang X et al. (2020) TGF-β inhibition via CRISPR promotes the long-term efficacy of CAR T cells against solid tumors. JCI insight 5 (4). doi:10.1172/jci.insight.133977
5. Wang Y, Tong C, Dai H et al. (2021) Low-dose decitabine priming endows CAR T cells with enhanced and persistent antitumour potential via epigenetic reprogramming. Nature Communications 12 (1):409. doi:10.1038/s41467-020-20696-x
6. Barber-Axthelm IM, Barber-Axthelm V, Sze KY et al. (2021) Stem cell–derived CAR T cells traffic to HIV reservoirs in macaques. JCI insight 6 (1). doi:10.1172/jci.insight.141502