Development and Utilization of Splice-specific Antibodies


Year of Award:
2019
Status:
Active
Award Type:
R21
Project Number:
CA240199
RFA Number:
RFA-CA-18-002
Technology Track:
Molecular & Cellular Analysis Technologies
PI/Project Leader:
O'NEILL, RACHEL
Other PI or Project Leader:
Not Applicable
Institution:
UNIVERSITY OF CONNECTICUT STORRS
ABSTRACTIn the human genome, alternative splicing events facilitate the generation of a proteome with agreater diversity than is observed in the protein-coding gene repertoire. In other words, the vastmajority of human genes can each produce numerous different transcripts and subsequently multipleprotein isoforms. However, aberrant splicing can produce an increase in novel isoforms or normallylow-level isoforms leading to potentially detrimental effects including altered protein function, protein-protein interactions as well as remodeling of protein complexes and pathways. In fact, the increasein these alternatively-spliced isoforms is now recognized as a major contributor to oncogenicphenotypes, such as the development of tumors and new blood vessels by supporting cell invasionand proliferation. A central challenge to realizing the biological impact, prognostic and therapeuticpotential of alternatively-spliced isoforms across multiple cancer types has been a lack of technologyavailable for unambiguously differentiating highly similar protein variants in their native forms. Currentmethods of detecting and studying alternatively-spliced isoforms at the protein level rely solely onindirect methods such as tag-based detection and nonspecific affinity reagents that are unable todiscriminate among multiple protein isoforms. Herein, we propose a novel approach to develop highlyspecific antibodies against alternatively-spliced protein isoforms using a targeting method at thesplice site junction with near amino acid specificity. With this technology, antibodies can be generatedto specifically detect a desired, alternatively-spliced protein isoform without cross-reactivity to the nativefull-length form. This technological advance will accelerate oncology research and enable scientiststo study both high-value and novel protein isoforms and their direct role in specific cancerphenotypes, metastatic potential, tumor grade specificity and survival rates, and provide a validationmeans for assessing the therapeutic potential of anticancer alternative-splicing inhibitors.