Constitutive oxalate-biodegrading Bacillus subtilis for kidney stones

用于肾结石的组成型草酸盐生物降解枯草芽孢杆菌

• 批准号: 10740242
• 负责人: Christoph Geisler
• 金额: $ 5.5万
• 依托单位: UNLOCKED LABS INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10740242
• 关键词: Adverse effects; Bacillus subtilis; Bacteria; Biodegradation; Calcium Oxalate; Circulation; Development; Diet; Distress; Economic Burden; Engineering; Enzymes; Hypokalemia; Kidney Calculi; Life; Microbe; Modeling; Nephrolithiasis; Oxalates; Pain; Patients; Persons; Pharmaceutical Preparations; Phase; Probiotics; Rattus; Recommendation; Recurrence; Reproduction spores; Small Business Innovation Research Grant; Testing; Thiazide Diuretics; Urine; Work; cost; design; effective therapy; experience; innovation; novel; overexpression; potassium citrate; side effect; small molecule; urinary

Project Summary / Abstract Kidney stones (nephrolithiasis) are an extremely painful and debilitating condition. About 12% of people will experience kidney stones in their lifetime, and about 70% of those will have recurrences. The economic burden of kidney stones is estimated to exceed $5 billion per year in the USA alone. Each year, over 3 million workdays are lost in the USA due to kidney stones. 75% of kidney stones consist of calcium oxalate (CaOx), and are caused by excessive urinary oxalate levels. Unfortunately, current treatment options all have serious drawbacks. A low oxalate diet is routinely recommended for CaOx stone patients, but is not very effective by itself. Alkalinizing agents such as potassium citrate can reduce stone recurrence, but are costly and often cause GI distress. Thiazide diuretics come with frequent, potentially life-threatening adverse effects such as hypokalemia. Thus, there remains a strong need for the development of new kidney stone treatment options that are both safe and effective. We propose to develop and test a new kidney stone treatment consisting of a novel Bacillus subtilis strain that overexpresses endogenous oxalate biodegrading enzymes. As bacteria stay in the gut, the spectrum of potential side effects is massively reduced as compared to small molecule drugs. Thus, our innovative approach is designed to result in an affordable, safe, and broadly effective kidney stone treatment option. In this Phase I SBIR project, we will develop new Bacillus subtilis strains engineered to biodegrade oxalate. We will produce and test different strains to optimize oxalate biodegradation capacity. As we will chromosomally overexpress endogenous enzymes in a spore-forming bacterium, our approach avoids key drawbacks of previously developed oxalate- degrading microbes. Finally, we will determine if the best performing engineered strain can effectively reduce urine oxalate levels in a relevant rat model.

项目概要/摘要 肾结石（肾结石）是一种极其痛苦且使人衰弱的疾病。关于 12%的人一生中会经历肾结石，其中约70%会患有肾结石 复发。肾结石造成的经济负担估计每年超过 50 亿美元 仅在美国。在美国，每年有超过 300 万个工作日因肾结石而损失。 75％的肾结石由草酸钙（CaOx）组成，是由于尿量过多引起的 草酸盐水平。 不幸的是，当前的治疗方案都有严重的缺点。低草酸盐饮食是 常规推荐用于氧化钙结石患者，但其本身并不是很有效。碱化 柠檬酸钾等药物可以减少结石复发，但价格昂贵且经常导致结石复发。 胃肠道不适。噻嗪类利尿剂经常出现可能危及生命的不良反应 如低钾血症。因此，仍然强烈需要开发新的肾脏 安全有效的石材治疗方案。 我们建议开发和测试一种新的肾结石治疗方法，其中包括一种新型的 过度表达内源草酸生物降解酶的枯草芽孢杆菌菌株。作为 细菌留在肠道内，潜在副作用的范围大大减少 与小分子药物相比。因此，我们的创新方法旨在实现 负担得起、安全且广泛有效的肾结石治疗选择。 在这个第一阶段 SBIR 项目中，我们将开发新的枯草芽孢杆菌菌株，经过改造， 生物降解草酸盐。我们将生产和测试不同的菌株以优化草酸盐 生物降解能力。因为我们将在染色体上过度表达内源酶 孢子形成细菌，我们的方法避免了先前开发的草酸盐的主要缺点 降解微生物。最后，我们将确定性能最佳的工程菌株是否可以 有效降低相关大鼠模型中的尿草酸盐水平。