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种新型的肌内基因递送平台，支持任何治疗性蛋白质的持续表达，这一能力尚未商业化。通过提供绕过当前实体瘤免疫疗法治疗的解决方案，预期提供双特异性自然杀伤细胞趋化因子（BiKE）治疗剂的持续表达和内源性分泌的能力将是范式转变。这种方法比现有技术侵入性更小，可以消除对治疗剂连续/频繁重复输注的需要，并且可以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，可能将治疗成本降低数万至数十万美元，并且适用于低资源环境，显着增加治疗的可用性。概念验证治疗将针对肝细胞癌，这是一种占肝癌90%的实体瘤。该平台具有广泛的应用，支持任何基因治疗应用（例如，单基因疾病），包括双特异性抗体T细胞增殖剂、治疗性抗体和候选疫苗（例如，内源性治疗性抗体的产生、DNA疫苗的递送和治疗性蛋白质的表达以治疗单基因罕见疾病）。该平台的预期影响包括提高治疗效果和改善患者生活质量。 该项目旨在推进安全，有效的肌内基因递送系统的开发，用于可重复的基因递送，以及证明该平台在体内表达内源性分泌的双特异性自然杀伤细胞因子（BIKE）的能力，用于治疗肝细胞癌（HCC），一种实体瘤。迄今为止，癌症治疗的基因治疗方法成本高、劳动密集且疗效有限。与注射裸DNA相比，该平台有望将基因递送提高1,000倍以上，并使基因产物能够有效分泌到血流中，从而允许系统表达。具体的目标是建立一个细胞表达系统的生产，纯化和功能验证的重组BiKE在体外，并使生物发光肝癌细胞系转导与商业化的慢病毒共表达RedFLuc和分泌GLuc更敏感的检测肿瘤的生存。 该项目还将验证BiKE表达构建体在人源化原位肝细胞癌（HCC）小鼠模型中的功效。将通过证明血清水平≥100-500 ng/ml时分泌型BiKE的持续全身表达≥ 1个月来确立概念验证，如通过酶联免疫吸附测定（ELISA）测定在施用后第3-60天评价的，该奖项反映了NSF的法定使命，并已被认为是值得通过评估使用基金会的智力价值和更广泛的支持影响审查标准。

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.