Engineering probiotic yeast for efficient sulforaphane delivery

工程益生菌酵母可有效输送萝卜硫素

• 批准号: 10305158
• 负责人: Charles Denby
• 金额: $ 5.2万
• 依托单位: BERKELEY FERMENTATION SCIENCE INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10305158
• 关键词: Anabolism; Animal Model; Biochemical; Biological Availability; Blood Circulation; Broccoli - dietary; Cauliflower; Cell Culture Techniques; Clinical Trials; Consumption; Development; Diffuse; Engineered Probiotics; Engineering; Enzymes; Gastrointestinal tract structure; Genetic; Goals; Health Benefit; Human; In Vitro; Isothiocyanates; Kale - dietary; Libraries; Malignant Neoplasms; Malignant neoplasm of prostate; Methods; Microbe; Mission; Myrosinase; Natural Products; Pathway interactions; Peripheral; Pharmaceutical Preparations; Phase; Plants; Preventive; Probiotics; Production; Property; Prostate Cancer therapy; Regimen; Research; Risk; Safety; Secretin; Signal Transduction; Small Business Innovation Research Grant; Source; Study models; Sulforaphane; Testing; Therapeutic; Tissues; United States National Institutes of Health; Vegetables; Yeasts; cancer prevention; cancer therapy; cruciferous vegetable; experimental study; food preparation; gastrointestinal; microbiome composition; mouse model; promoter; sugar

Project Summary/Abstract Consumption of cruciferous vegetables such as broccoli, brussels sprouts, and cauliflower has been shown to reduce the risk of developing multiple forms of cancer. Recent research has suggested that these health benefits result in part from the biochemical activity of isothiocyanates, a class of natural products found in these vegetables. Sulflurophane (SFN), one such isothiocyanate produced in abundance by crucifers like broccoli and kale, has shown especially impressive potential as a chemo-preventative and cancer therapeutic in in vitro and animal model studies. It has been challenging to rigorously assess the health benefits of SFN in humans, however. These challenges result from SFN’s inherent instability, as well as the varying effects that different food preparation methods and personal microbiome compositions have on SFN bioavailability. In this SBIR, we propose to engineer a probiotic yeast that will biosynthesize SFN from within the human gastrointestinal-tract. This probiotic yeast will provide a consistent and direct source of SFN that readily diffuses into the bloodstream, and is not dependent on specific microbiome compositions, food preparation methods, or drug regimens. Given the impressive body of research detailing the potential for SFN to treat prostate cancer in cell culture and animal models, we will develop this probiotic for potential use as a prostate cancer therapeutic. The probiotic microbe we will use as a host for our engineering efforts is the safe, and genetically tractable probiotic yeast, S. boulardii. The engineered S. boulardii strain we develop will provide a means to directly assess the health benefits of SFN in prostate cancer therapy, while also providing a mechanism to efficiently deliver SFN for cancer therapeutic and preventative purposes. Phase I experiments will develop an S. boulardii strain that secretes a highly active myrosinase enzyme that is able to perform the terminal step in SFN biosynthesis. To achieve this goal, we will screen libraries of myrosinases, secretin signals, and yeast promoters to identify the genetic components that most effectively produce SFN under gastrointestinal conditions. To establish a proof-of-principle and motivate further research, Phase I will seek to engineer S. boulardii for production of 200umoles SFN per day, as small clinical trials that administered this quantity have shown encouraging results. In Phase II research, we will further engineer S. boulardii such that it contains the entire SFN biosynthesis pathway and produces SFN from fermentable sugars. Resultant strains will be tested in prostate cancer mouse models for safety and cancer-preventive properties. This project is directly in line with the NIH mission, as it will advance our understanding of a promising cancer therapeutic molecule, while also providing a direct means of delivering this potential therapeutic for cancer therapy and prevention.

项目总结/摘要 食用十字花科蔬菜，如花椰菜、布鲁塞尔子甘蓝和花椰菜， 降低患多种癌症的风险。最近的研究表明，这些健康 这些益处部分来自异硫氰酸酯的生物化学活性，异硫氰酸酯是一类天然产物， 这些蔬菜。硫氟凡（SFN），一种由十字花科植物（如 西兰花和羽衣甘蓝作为化学预防和癌症治疗的潜力特别令人印象深刻 在体外和动物模型研究中。严格评估SFN的健康益处一直具有挑战性， 然而，人类。这些挑战源于SFN固有的不稳定性，以及 不同的食物制备方法和个人微生物组组成对SFN生物利用度有影响。在这 SBIR，我们建议工程益生菌酵母，将生物合成SFN从人体内 胃肠道这种益生菌酵母将提供一种稳定和直接的SFN来源， 扩散到血液中，并且不依赖于特定的微生物组组成， 方法或药物方案。鉴于令人印象深刻的研究机构详细说明了SFN治疗的潜力 在细胞培养和动物模型中研究前列腺癌，我们将开发这种益生菌作为前列腺的潜在用途 癌症治疗我们将使用的益生菌作为我们工程工作的宿主是安全的， 遗传上易处理的益生菌酵母，S.布拉地。经过改造的S.我们开发的布拉氏菌菌株将提供一种 意味着直接评估SFN在前列腺癌治疗中的健康益处，同时还提供 用于癌症治疗和预防目的的有效递送SFN的机制。I期实验 会发展出一个S。布拉氏菌菌株，分泌一种高活性黑芥子酶，能够执行 SFN生物合成的末端步骤。为了实现这一目标，我们将筛选黑芥子酶、分泌素 信号和酵母启动子，以鉴定最有效地产生SFN的遗传组分。 胃肠道疾病为了建立一个原理证明并推动进一步的研究，第一阶段将寻求 工程师S.布拉氏菌每天生产200 umoles SFN，作为小型临床试验， 数量已经显示出令人鼓舞的结果。在第二阶段的研究中，我们将进一步工程S。布拉地，这样它 包含整个SFN生物合成途径并从可发酵糖产生SFN。所得菌株 将在前列腺癌小鼠模型中测试其安全性和癌症预防特性。这个项目是 直接符合NIH的使命，因为它将促进我们对一种有前途的癌症治疗方法的理解 分子，同时还提供了递送这种用于癌症治疗的潜在治疗剂的直接手段， 预防