SBIR Phase I: Endogenously secreted bispecific natural killer cell engagers (BIKEs) for therapy of solid tumors

这个小型企业创新研究(SBIR)第一阶段项目提供了一个新颖的肌肉内基因传递平台，支持任何治疗性蛋白质的持续表达，这一能力尚未商业化实现。通过提供一种绕过目前实体肿瘤免疫治疗的解决方案，提供双特异性自然杀伤细胞激活因子(Bike)治疗药物的持续表达和内源性分泌的能力有望转变为范式。这种方法比目前最先进的方法侵入性更小，可以消除持续/频繁地重复输入治疗药物的需要，并将避免在给药期间住院的需要。该方法具有较低的价格点，与其他免疫疗法相比，有可能将治疗成本降低数万至数十万美元，并且能够适应低资源环境，显著增加治疗的可获得性。这项概念验证疗法将针对肝细胞癌，这是一种实体肿瘤，占肝癌的90%。该平台具有广泛的应用，支持交付任何可以受益于分泌蛋白系统表达的基因治疗应用(例如，单基因疾病)，包括双特异性抗体T细胞结合蛋白、治疗性抗体和候选疫苗(例如，内源性治疗性抗体生产、DNA疫苗的交付以及治疗单基因罕见疾病的治疗性蛋白的表达)。该平台的预期影响包括改善治疗效果和改善患者的生活质量。该项目旨在促进一种安全、高效的肌肉内基因传递系统的开发，用于可编辑的基因传递，并展示该平台在体内表达内源性分泌的双特异性自然杀伤细胞活体(Bikes)用于治疗实体瘤肝细胞癌(HCC)的能力。到目前为止，用于癌症治疗的基因治疗方法一直是昂贵的、劳动密集型的，而且疗效有限。与注射裸DNA相比，这一平台有望将基因传递提高1000倍以上，并使基因产品能够有效地分泌到血液中，从而实现系统表达。其具体目的是建立一个细胞表达系统，用于体外生产、纯化和功能验证，并使转导商品化慢病毒的生物发光肝癌细胞系共表达RedFLuc和分泌型GLuc，以更敏感地检测肿瘤存活。该项目还将在人源化的原位肝细胞癌(HCC)小鼠模型中验证Bike表达构建的有效性。概念验证将建立在≥持续系统表达1个月的≥血清水平100-500 ng/ml，在肌肉注射后3-60天通过酶联免疫吸附分析(ELISA)分析进行评估。该奖项反映了国家科学基金会的法定使命，并已通过使用基金会的智力价值和更广泛的影响审查标准进行评估而被认为值得支持。

This Small Business Innovation Research (SBIR) Phase I project provides a novel, intramuscular gene delivery platform that supports sustained expression of any therapeutic protein, a capability yet to be commercially realized. The ability to provide sustained expression and endogenous secretion of bispecific natural killer cell engager (BiKE) therapeutics is expected to be paradigm shifting by providing a solution that bypasses current immunotherapy treatments for solid tumors. This approach, which is less invasive than the current state-of-the art, could eliminate the need for continuous/frequent repeated infusions of therapeutics and would circumvent the need for hospitalization during administration. The approach has a lower price point, potentially reducing the cost of therapy by tens to hundreds of thousands of dollars compared to other immunotherapies, and is amenable to low resource settings, significantly increasing the availability of treatment. The proof-of-concept therapeutic will target hepatocellular carcinoma, a solid tumor that accounts for 90% of liver cancers. The platform has broad applications, supporting delivery of any gene therapy application (e.g., monogenic disease) that can benefit from systemic expression of a secreted protein, including bi-specific antibody T cell engagers, therapeutic antibodies, and vaccine candidates (e.g., endogenous therapeutic antibody production, delivery of DNA vaccines, and expression of therapeutic proteins to treat monogenic rare diseases). Anticipated impacts of the platform include improved treatment efficacy and improved patient quality of life. This project seeks to advance the development of a safe, efficient intramuscular gene delivery system for redosable gene delivery as well as the demonstration of the platform’s ability to express endogenously secreted bispecific natural killer cell engagers (BIKEs) in vivo for treatment of hepatocellular carcinoma (HCC), a solid tumor. To date, gene therapy approaches to cancer treatment have been costly, labor intensive, and limited in efficacy. This platform is expected to enhance gene delivery by over 1,000-fold compared to the injection of naked DNA and to enable efficient secretion of the gene product into the blood stream, thereby allowing for systemic expression. Specific aims are to establish a cell expression system for production, purification, and functional validation of the recombinant BiKE in vitro and to make bioluminescent hepatoma cell lines transduced with a commercialized lentivirus co-expressing RedFLuc and secreted GLuc for more sensitive detection of tumor survival. The project will also validate the efficacy of the BiKE expression construct in a humanized, orthotopic hepatocellular carcinoma (HCC) mouse model. Proof of concept will be established with the demonstration of sustained systemic expression of the secretable BiKE for ≥ 1 month at serum levels of ≥100-500 ng/ml, as evaluated by enzyme-linked immunosorbent assay (ELISA) assays at days 3-60 post-intramuscular delivery.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.