A membrane-based immunotherapy for triple negative breast cancer

描述(申请人提供)：三阴性乳腺癌(TNBC)，包括缺乏雌激素、孕激素和HER-2受体蛋白的肿瘤，占美国所有乳腺癌诊断的15%-20%，是肿瘤切除后开发有效治疗方法最具挑战性的癌症之一。即使采用传统的放疗和化疗方案，患者的预后也很差，与其他乳腺癌相比，他们经历了早期、频繁的复发。迫切需要新的治疗方法。我们建议开发和测试一种治疗转移性TNBC的免疫疗法，使用一种个性化的、容易实现的方法，并且不需要建立细胞系或基因转移。这种专有的癌症免疫治疗技术的基本原理是将糖脂锚定形式的免疫刺激分子(GPI-ISM)，如B7-1和IL-12，结合到来自手术切除的患者组织的肿瘤膜小泡(TMV)上。我们期望这些附着在膜上的免疫刺激分子能同时传递肿瘤抗原并激活免疫细胞，从而促进有效的抗肿瘤免疫反应。提议的疫苗的新颖性不在于免疫刺激分子的使用，而在于它们的使用方式。蛋白质转移方法与以前使用基因转移技术或分泌型细胞因子开发癌症疫苗的研究有很大不同。主要优点是免疫治疗的准备工作可以在短短几天内完成。此外，肿瘤膜制剂可以作为冷冻等份储存，并为加强注射而生物库。这些因素将大大降低免疫疗法的生产和治疗成本。在这一阶段的应用中，我们将使用这种蛋白质转移技术进行基于膜的乳腺癌免疫治疗，并在小鼠模型中测试其抑制三重阴性乳腺癌转移的效果。具体地说，我们将：1)制造GPI-ISM，优化TMV的加工和蛋白质转移，并使用小鼠转移的TNBC模型评估疗效；2)(A)确定基于膜的免疫疗法是否可以预防/减少TNBC的转移。(B)调查联合使用抗免疫抑制剂是否提高了基于膜的TNBC免疫疗法的疗效。完成拟议的SBIR第一阶段可行性研究将证明基于膜的免疫疗法在抑制三阴性乳腺癌转移生长方面的有效性。这将为第二阶段的研究铺平道路，如毒性和规模化的GMP/GLP条件，以大规模生产纯化的人GPI-分子，这将有助于我们将疫苗产品推进到人体临床试验。

DESCRIPTION (provided by applicant): Triple negative breast cancer (TNBC), which includes tumors lacking estrogen, progesterone, and HER-2 receptor proteins, represents 15-20% of all breast cancer diagnoses in the US, and is one of the most challenging cancers for developing an effective therapy post tumor resection. Even with conventional radiation and chemotherapy regimens, patients have poor prognosis, experiencing early, frequent relapses in comparison to other breast cancers. New therapies are critically needed. We propose to develop and test an immunotherapy to treat metastatic TNBC using an approach that is personalized, easy to accomplish, and not requiring establishment of cell lines or gene transfer. The defining principle of this proprietary cancer immunotherapy technology is the incorporation of glycolipid anchored forms of immunostimulatory molecules (GPI-ISMs), such as B7-1 and IL-12, onto tumor membrane vesicles (TMV) derived from surgically removed patient tissue. We expect that these membrane attached immunostimulatory molecules will simultaneously deliver tumor antigens and activate immune cells to promote an effective anti-tumor immune response. The novelty of the proposed vaccine lies not in the use of immunostimulatory molecules but in the way they are employed. The protein transfer approach is substantially different from previous studies that used gene transfer techniques or secretory cytokines to develop cancer vaccines. The major advantage is that the immunotherapy preparation can be accomplished within just a few days. Additionally, tumor membrane preparations can be stored as frozen aliquots and biobanked for booster injections. These factors will significantly reduce the cost of immunotherapy production and treatment. In this Phase I application, we will use this protein transfer technique for membrane-based breast cancer immunotherapy and test its efficacy in inhibiting triple negative breast cancer metastasis in a murine model. Specifically, we will: 1) manufacture GPI-ISMs, optimize TMV processing and protein transfer, and evaluate efficacy using a murine metastatic TNBC model, 2) (A) Determine whether the membrane-based immunotherapy prevents/reduces metastasis of TNBC. (B) Investigate whether co-administration of anti-immunosuppressive agents augments efficacy of the membrane- based TNBC immunotherapy. Completion of the proposed SBIR Phase I feasibility studies will demonstrate the efficacy of membrane-based immunotherapy in inhibiting the metastatic growth of triple negative breast cancers. This will pave the way for Phase II studies such as toxicity and standardization of GMP/GLP conditions for large-scale production of purified human GPI-molecules, which will help us advance the vaccine product to human clinical trials.