PET Imaging of TIGIT Expression on Tumor-Infiltrating Lymphocytes.

Abstract

Therapeutic checkpoint inhibitors on tumor-infiltrating lymphocytes (TIL) are being increasingly utilized in the clinic. The T-cell immunoreceptor with Ig and ITIM domains (TIGIT) is an inhibitory receptor expressed on T and natural killer cells. The TIGIT signaling pathway is an alternative target for checkpoint blockade to current PD-1/CTLA-4 strategies. Elevated TIGIT expression in the tumor microenvironment correlates with better therapeutic responses to anti-TIGIT therapies in preclinical models. Therefore, quantifying TIGIT expression in tumors is necessary for determining whether a patient may respond to anti-TIGIT therapy. PET imaging of TIGIT expression on TILs can therefore aid diagnosis and in monitoring therapeutic responses. Antibody-based TIGIT imaging radiotracers were developed with the PET radionuclides copper-64 (⁶⁴Cu) and zirconium-89 (⁸⁹Zr). <i>In vitro</i> characterization of the imaging probes was followed by <i>in vivo</i> evaluation in both xenografts and syngeneic tumor models in mouse. Two anti-TIGIT probes were developed and exhibited immunoreactivity of >72%, serum stability of >95%, and specificity for TIGIT with both mouse TIGIT-expressing HeLa cells and <i>ex vivo</i>-activated primary splenocytes. <i>In vivo</i>, the ⁸⁹Zr-labeled probe demonstrated superior contrast than the ⁶⁴Cu probe due to ⁸⁹Zr's longer half-life matching the TIGIT antibody's pharmacokinetics. The ⁸⁹Zr probe was used to quantify TIGIT expression on TILs in B16 melanoma in immunocompetent mice and confirmed by <i>ex vivo</i> flow cytometry. This study develops and validates novel TIGIT-specific ⁶⁴Cu and ⁸⁹Zr PET probes for quantifying TIGIT expression on TILs for diagnosis of patient selection for anti-TIGIT therapies.