Monitoring Treatment Efficacy in Leprosy

项目摘要 麻风病是由麻风分枝杆菌引起的，麻风分枝杆菌是一种生长缓慢的细菌，不能在实验室中培养 媒体这种疾病是可以治愈的，早期使用多种药物可以预防残疾 治疗（MDT）。MDT的实施成功地减少了报告的新病例数， 1981年为1000万，2005年为25万，但此后一直保持稳定，2019年新增病例为20万，这表明 疾病传播仍然活跃一个重要的因素要考虑在持续传播的 麻风病是治疗效果。在临床实践中，治疗有效性的主观评估是基于 在6至12个月长的治疗结束时对皮肤病变进行临床检查， 患者可能会终身残疾。复发在地方性环境中很常见，但可能仅 在完成治疗后10至15年出现临床症状，这代表了对 后续研究，并使新的药物方案的临床试验极具挑战性。因此存在 明确需要简单的方法，快速准确地评估治疗效果，直接从临床 在化疗期间而不是在治疗结束时采集样本。抗麻风药物的筛选与优化 具有缩短治疗持续时间潜力的药物和药物组合方案同样复杂 事实上，M。麻风不能在体外培养。 目前测量临床和临床前样品中细菌活力的金标准方法包括： 将从这些样品制备的细菌接种到免疫受损小鼠的足垫中 随后在感染后6至12个月对脚垫中的细菌复制进行显微镜评估。 除了耗时之外，这种方法还昂贵，技术上具有挑战性，并且需要维护 在BSL 2或BSL 3动物设施中的几个月的大量小鼠。 在这个项目中，我们建议评估一些分子方法，包括最近的一个 内部开发，以快速监测对化疗治疗的反应， 样品（目标1）和M.麻风感染（目的2）。 这一项目的成功可能对监测麻风病人的治疗反应产生变革性影响， 使临床医生能够“实时”评估治疗成功或失败。分子读数显示 还有望提供急需的方法来快速评价治疗效果， 这些模型和人体模型可用于筛选和优化新药和药物制剂。 联合方案。

Project Summary Leprosy is caused by Mycobacterium leprae, a slow-growing bacterium that cannot be cultured in laboratory media. The disease is curable, and disabilities can be prevented with the early administration of multi-drug therapy (MDT). The implementation of MDT successfully decreased the number of new cases reported from 10 million in 1981 to 250,000 in 2005 but plateaued since then with 200,000 new cases in 2019, suggesting that disease transmission is still active. One important factor to consider in the sustained transmission of leprosy is treatment efficacy. In clinical practice, treatment effectiveness is subjectively assessed based on clinical examination of skin lesions at the end of the 6 to 12-month-long treatment, a long period of time during which patients may develop permanent disabilities. Relapse is common in endemic settings but might only manifest clinically 10 to 15 years after completion of therapy which represents a considerable obstacle to follow-up studies and renders the clinical testing of new drug regimens extremely challenging. There is thus a clear need for easy methods that rapidly and accurately assesses treatment efficacy directly from clinical samples during chemotherapy rather than at the end of treatment. Screening and optimization of anti-leprosy drugs and drug combination regimens with the potential to shorten treatment duration is similarly complicated by the fact that M. leprae cannot be cultured in vitro. The current gold standard method to measure bacterial viability in clinical and pre-clinical samples involves the inoculation of bacteria prepared from these samples into the footpads of immunocompromised mice followed by the microscopic assessment of bacterial replication in the footpads 6 to 12 months post-infection. Besides being time-consuming, this method is expensive, technically challenging, and requires maintaining a high number of mice for several months in BSL2 or BSL3 animal facilities. In this project, we propose to evaluate a number of molecular methods, including one recently developed in-house, to rapidly monitor response to chemotherapeutic treatments using both human samples (Aim 1) and an animal model of M. leprae infection (Aim 2). Success in this project could be transformative in monitoring treatment response in leprosy patients by enabling clinicians to assess in “real-time” treatment success or failure. The molecular read-outs developed are further expected to provide much-needed methods to rapidly evaluate treatment efficacy both in animal models and in humans that may be used in the screening and optimization of new drugs and drug combination regimens.