A microbiome-informed platform for the development and testing of bacterial therapies for colorectal cancer

项目摘要 有一个明确的当务之急是开发有效的，具有成本效益的治疗方法，以对抗 癌症给社会带来的挑战在这里，我们通过综合开发来满足这一需求。 工程细胞对广泛的癌症类型有效，特别强调 结直肠癌（CRC）。这种癌症类型是癌症死亡的第二大常见原因 在美国，每年有超过5万美国人死亡。最近的研究表明， 基因工程的力量，使更有效的癌症的重大进展， 疗法基因工程细胞的引入，如嵌合抗原受体T (CAR T）细胞，已经显示出治疗许多类型的B细胞恶性肿瘤的巨大前景，但是 不幸的是，将CAR T细胞靶向实体瘤仍然具有挑战性。在这个项目中，我们将 利用合成生物学的工具， 而不是哺乳动物细胞。某些细菌物种已经证明了一种有用的“归巢”能力。 并且选择性地定殖实体肿瘤而不感染健康组织。这种肿瘤靶向 财产将被利用在拟议的工作，提供安全，有效的治疗直接向 最需要它们的位置：肿瘤的实核。以前我们开发了一个 细菌治疗，并在转移性疾病的动物模型中进行了测试。相对于其他 利用细菌细胞的方法，这种“裂解菌株”不需要专门的遗传学。 当编码货物的分泌发生改变时，它只是将其释放到环境中， 细胞就会破裂最初，我们将对裂解菌株进行遗传修饰，以产生广泛的 用于测试的治疗剂，包括毒素（来自细菌、动物和植物），酶， 抗生素和凋亡肽。接下来，我们将分析来自人类的肿瘤微生物组。 因为我们假设天然细菌种群的组成将提供一个 独特的签名（类似于指纹），可用于将肿瘤分为不同的 亚型我们期望利用这些指纹来鉴定其他具有上级适应性的物种 用于治疗CRC的治疗性递送。一旦确定，我们将开发两种体外试验， 测试候选菌株。我们将使用微流控技术来创造一个高通量的 用于细菌和癌细胞系的共培养系统。与此同时，我们将开发一种共同培养 用于从相同的人类肿瘤样品产生的细菌和类器官的系统 该技术以前曾用于菌株鉴定和指纹图谱。最后，我们将测试 最有希望的治疗方法，以确定在结肠直肠癌的动物模型中的疗效， 临床模型

Project Summary There is a clear imperative to develop potent, cost effective therapeutics to confront the challenge cancer poses to society. Here we address this need by developing synthetically engineered cells effective against a broad range of cancer types with a special emphasis on colorectal cancer (CRC). This cancer type is the second most common cause of cancer death in the US, with more than 50,000 Americans dying every year. Recent research demonstrates the power of genetic engineering to make significant advances towards more efficacious cancer therapy. The introduction of genetically engineered cells, such as chimeric antigen receptor T (CAR T) cells, has shown great promise for treating many types of B cell malignancies, but unfortunately targeting CAR T cells to solid tumors remains challenging. In this project we will use the tools of synthetic biology to make new engineered therapies based on bacterial rather than mammalian cells. Certain bacterial species have demonstrated a useful ability to “home in” and selectively colonize solid tumors without infecting healthy tissue. This tumor targeting property will be exploited in the proposed work to deliver safe, effective therapies directly to the locations where they are needed most: the solid core of tumors. Previously we developed a bacterial therapeutic and tested it in an animal model of metastatic disease. In contrast to other approaches utilizing bacterial cells, this “lysis strain” does not require specialized genetic modifications for the secretion of encoded cargo, it simply releases it into the environment when the cells burst. Initially we will genetically modify the lysis strain to produce a wide range of therapeutics for testing, including toxins (from bacteria, animals and plants), enzymes, antibiotics, and apoptotic peptides. Next we will analyze the tumor microbiome from human samples since we hypothesize that the native bacterial population's composition will provide a unique signature (analogous to a fingerprint) that can be used to divide tumors into distinct subtypes. We expect to use these fingerprints to identify other species with superior suitability for therapeutic delivery in treating CRC. Once identified we will develop two in vitro assays for testing the candidate strains. We will use microfluidic technology to create a high throughput co-culturing system for bacteria and a cancer cell line. In parallel, we will develop a co-culturing system for bacteria and organoids that are generated from the same human tumor samples which had been previously used for strain identification and fingerprinting. Lastly we will test the most promising therapies in an animal model of colorectal cancer to determine efficacy in a pre- clinical model.