PROJECT ABSTRACT: The goal of this Phase II SBIR proposal is to further validate the genetically defined Oncopig hepatocellular carcinoma (HCC) model—capable of modeling diverse HCC driver mutational profiles through induced KRASG12D and TP53R167H expression and subsequent CRISPR editing—for preclinical evaluation of locally delivered, personalized HCC therapies. HCC is an aggressive liver malignancy representing the 7th most common cancer and the 4th most common cause of cancer death worldwide, illustrating the critical need for improved HCC treatment options. Since HCC and other cancers are driven by the accumulation of genetic driver mutations conferring selective growth advantages, personalized cancer models are required to evaluate targeted therapeutics for this deadly disease. Furthermore, the use of transarterial delivery-based approaches for HCC treatment combined with similarities in size and drug metabolism between pigs and humans highlights the critical need and translational value of the genetically defined Oncopig HCC model for investigating novel and re-purposed therapeutics targeting specific driver mutations. Importantly, there is broad interest in the use of Oncopigs in preclinical trials (see Support Letters). This proposal will demonstrate differential efficacy of transarterial targeted PI3K inhibitor delivery to Oncopig PTENKO (PI3K inhibitor responsive) and KEAP1KO (PI3K inhibitor non-responsive) HCC tumors using a clinically relevant trial design. Use of liquid biopsies for minimally invasive HCC driver mutation quantification (see Volition Support Letter) will facilitate translation of HCC precision medicine approaches into clinical practice, currently lacking due to infrequency of routine HCC biopsy collection for biological profiling. The genetically defined Oncopig HCC model will be validated for targeted therapeutic testing by pursuing the following Specific Aims: (1) Characterize genetically defined Oncopig HCC tumors over a clinically relevant 3-month diagnostic monitoring period. (2) Demonstrate delivery of targeted PI3K inhibitor therapeutic doses to Oncopig HCC tumors via transarterial embolization. (3) Demonstrate efficacy of transarterial targeted PI3K inhibitor delivery for Oncopig PTENKO HCC in a clinically relevant 3-month follow-up timeframe. The ability to achieve quantifiable therapeutic PI3K inhibitor concentrations following transarterial delivery in the genetically defined Oncopig HCC model will be demonstrated, in addition to a clinically relevant difference in response (40% increase in complete response rate in PTENKO compared to KEAP1KO HCC) based on clinically employed mRECIST guidelines. This work will validate the genetically defined Oncopig HCC model for targeted therapeutic evaluation, enabling Sus Clinicals to provide personalized tumor development and targeted therapeutic testing services using the Oncopig platform. Future work focused on expanding Sus Clinicals commercialization of this techno...