Pre-clinical testing of low intensity ultrasound as novel strategy to prevent paclitaxel-induced hair follicle damage in a humanized mouse model of chemotherapy-induced alopecia

项目摘要 紫杉烷类化合物如紫杉醇（PTX）是一种高效的抗微管药物，常用于治疗癌症 这些药物虽然不能用于治疗，但也会引起主要的剂量限制性皮肤毒性。其中最可怕的是脱发， 甚至是永久性的。PTX诱导的脱发（PIA）严重影响患者的生活质量，并可能导致 拒绝挽救生命的化疗虽然头皮脱发可能会减少头皮冷却，这项技术是 尚未广泛使用，不能应用于眉毛，睫毛和面部毛发， 效益因此，预防这种急性和慢性癌症相关的发病率不仅可以减少痛苦， 与PIA相关的焦虑和抑郁，但也可能改善药物依从性。 为此，我们的项目旨在为保护头发的创新策略提供原理证明 低强度超声（LIUS）是一种广泛使用的 医疗技术，安全性极佳。我们已经证明，PTX稳定微管， 高度增殖的毛基质角质细胞，从而诱导其凋亡并引起脱发。此外，本发明还提供了一种方法， PTX还诱导主要的HF干细胞损伤，这可以消除HF的再生能力。我们有 还发现，短暂暴露于LIUS可以有效地中和PTX对培养细胞的细胞毒性作用。 细胞通过破坏PTX诱导的刚性微管束，从而防止细胞死亡。最重要的是我们 已经产生了初步的证据，LIUS也保护器官培养的人类头皮HFs及其 离体上皮干细胞以及体内小鼠HF免于PTX毒性。最后，我们建立了一个 通过用以下物质处理SCID/米色小鼠上的人头皮皮肤异种移植物来研究PIA的人源化小鼠模型 PTX。这使我们能够第一次在体内研究候选PTX保护性干预措施， 最佳地模拟人类PIA的临床现实的条件。 作为将这种PIA预防策略引入临床的关键一步，我们建议测试 使用我们的人源化PIA小鼠模型，在临床前确定LIUS是否在体内也具有PIA保护性。具体地说， 我们将确定LIUS如何影响人类头发基质的微管网络、功能和存活 角质形成细胞和HF干细胞的急性和重复PTX治疗。这些研究将揭示 LIUS在体内对PTX对人HF的急性和慢性损伤提供保护。预期 结果将指导后续设计临床试验，探讨LIUS在临床PIA中的疗效 卵巢癌的治疗方法如果成功，这种创新的，无毒的，容易翻译的， 广泛可用的，经济的，非常好的耐受性PIA预防策略将大大提高质量， 许多紫杉烷治疗的癌症患者的生活，通过将他们从肿瘤药物的主要皮肤毒性中解放出来， 治疗，从而改善药物依从性。

PROJECT SUMMARY Taxanes like paclitaxel (PTX) are highly effective anti-microtubule agents frequently used in cancer therapy, but they also cause major dose-limiting skin toxicity. The most dreaded of these is hair loss, which can even be permanent. PTX-induced alopecia (PIA) severely impacts patients’ quality of life, and may lead to refusal of life-saving chemotherapy. While scalp hair loss may be reduced by scalp cooling, this technology is not yet widely available, cannot be applied to eyebrows, eyelashes and facial hair, and is of unpredictable benefit. Therefore, prevention of this acute and chronic cancer-related morbidity will not only reduce distress, anxiety, and depression associated with PIA, but likely also improve medication adherence. To this end, our project aims to generate proof-of-principle for the innovative strategy to protect hair follicles (HFs) from alopecia by applying low intensity ultrasound (LIUS), a much-used and widely available medical technology with an excellent safety profile. We have demonstrated that PTX stabilizes microtubules in highly proliferative hair matrix keratinocytes, thus inducing their apoptosis and causing hair loss. In addition, PTX also induces major HF stem cell damage, which can obliterate the HF’s capacity to regenerate. We have also discovered that a brief exposure to LIUS can effectively neutralize the cytotoxic effects of PTX on cultured cells by disrupting PTX-induced rigid microtubule bundles and thus prevent cell death. Most importantly, we have generated preliminary evidence that LIUS also protects organ-cultured human scalp HFs and their epithelial stem cells ex vivo as well as mouse HFs in vivo from PTX toxicity. Finally, we have established a humanized mouse model for studying PIA by treating human scalp skin xenografts on SCID/beige mice with PTX. This enables us, for the first time, to study candidate PTX-protective interventions under in vivo conditions that optimally mimic the clinical reality of human PIA. As a critical step towards introducing this PIA prevention strategy into the clinic, we propose to test preclinically whether LIUS is also PIA-protective in vivo, using our humanized PIA mouse model. Specifically, we will determine how LIUS impacts on the microtubule network, function, and survival of human hair matrix keratinocytes and HF stem cells under acute and repetitive PTX therapy. These studies will reveal whether LIUS provides protection against acute and chronic damage by PTX to human HFs in vivo. The expected results will guide the subsequent design of a clinical trial that probes the efficacy of LIUS in clinical PIA prevention during ovarian cancer management. If successful, this innovative, drug-free, easily translatable and widely available, economical, and very well-tolerated PIA prevention strategy will greatly improve the quality of life of numerous taxane-treated cancer patients by liberating them from a major skin toxicity of oncological therapy and will thus improve medication adherence.