Therapeutic strategies for treatment of giant congenital melanocytic nevi

巨大先天性黑素细胞痣的治疗策略

• 批准号: 10570507
• 负责人: DAVID E FISHER
• 金额: $ 51.17万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10570507
• 关键词: Ablation; Acids; Agonist; Anguish; Animal Model; Anti-Inflammatory Agents; Binding; Body Surface; CD8-Positive T-Lymphocytes; CD8B1 gene; Child; Childhood; Childhood Melanoma; Clinical; Clinical Trials; Complement; Congenital melanocytic nevus; Cutaneous; Deformity; Dermatology; Disease; Dose; Engineering; Esters; Excision; Exhibits; Funding; Genetic; Genetically Engineered Mouse; Goals; Haptens; Histology; Human; Hyperpigmentation; Imiquimod; Immunity; Immunodeficient Mouse; Immunologic Deficiency Syndromes; Individual; Infant; Inflammation; Inflammatory; Institutional Review Boards; Kinetics; Laser injury; Lasers; Lesion; LoxP-flanked allele; MEK inhibition; MEKs; Macrophage; Measurable; Medical; Melanocytic Neoplasm; Melanocytic nevus; Melanosomes; Methods; Modeling; Modification; Morbidity - disease rate; Mouse Strains; Mus; National Institute of Arthritis, and Musculoskeletal, and Skin Diseases; Natural Immunity; Natural Killer Cells; Neoplasms; Nevus; Nevus Cell; Oncogenes; Operative Surgical Procedures; PIK3CG gene; Peptides; Pharmacotherapy; Phase; Pre-Clinical Model; Prevention; Probability; Publishing; Regimen; Regressing Melanoma; Reporting; Reproducibility of Results; Resected; Risk; SILV gene; Schedule; Skin; Symptoms; T-Lymphocyte; Tamoxifen; Testing; Therapeutic; Tissues; Titrations; Topical application; Toxic effect; Transplantation; Treatment-related toxicity; Visual; Xenograft procedure; adaptive immunity; anti-PD-1; constitutive expression; effector T cell; giant congenital nevus; human disease; inducible Cre; inducible gene expression; inhibitor; kinase inhibitor; melanocyte; melanoma; mortality; mouse model; neoantigens; pre-clinical; prevent; promoter; recruit; senescence; skin lesion; treatment strategy; tumor

Abstract Congenital Melanocytic Nevi (CMN) can reach giant sizes, transform to childhood melanoma, and thus trigger pre-emptive surgery inducing profound morbidity. In the prior funding period, we constructed four CMN models to test regression-inducing treatments aimed at avoiding surgery. Three are genetically engineered mice driven by the (floxed)NRASQ61R oncogene, responsible for most human CMN, with melanocyte-targeting via tamoxifen- inducible or constitutive expression from Tyr or Dct promoters (varied strengths). The fourth model is xenografted resected human CMNs on immunodeficient mice. Several models displayed melanoma transformation after long latency. Inducible NRASQ61R clarified the kinetics of proliferative vs senescent nevus phases and verified regressive treatments to succeed in either phase. Tyr-Cre;NRASQ61R exhibited severe human-like CNS and skin lesions whereas the weaker Dct-Cre induced a more typical skin-restricted giant nevi for which inhibitors of MEK, PI3K, or cKIT induced incomplete (~80%) regressions and combinations fully depleted visual nevocytes. Topical treatment with the proinflammatory hapten Squaric Acid Dibutyl Ester (SADBE) induced complete visual CMN regression after 3 doses. This regimen also fully prevented murine melanoma transformation, within all treated nevus skin. We found that topical SADBE requires macrophages (not B, T, or NK cells), so it could be tested in human CMN xenografted onto SHO mice (which lack adaptive immunity but retain macrophages). SADBE recruited murine macrophages into the human xenografts and regressed ~90% of the human nevocytes. Since SADBE is already used elsewhere in dermatology, and has been reported to induce “depigmentation,” we plan to test it clinically. However, since clinical trials with efficacy endpoints are not supported in R01s from NIAMS, we are seeking separate funding to support this clinical trial that already received FDA/IND and IRB approvals. While we are excited by the models and strategies uncovered during this funding period, we believe it is crucial to build on this progress to increase the likelihood of achieving real therapeutic breakthrough for this devastating children’s disease. Accordingly Aims 1-3 use our models to boldly seek further enhanced efficacy and minimized toxicity of treatments: Aim 1 deeply and systematically develops kinase inhibitor combinations. It also tests MEK inhibitors (MEKi) with SADBE, as both exhibit individual efficacy and MEKi was recently shown to promote M1- like (pro-inflammatory) over M2-like macrophage differentiation, a mechanistic feature that may enhance SADBE’s efficacy. Aim 2 seeks localized T cell adaptive immunity recruitment, to complement SADBE’s macrophage induction. We have shown anti-PD1 triggers recruitment of melanocyte (gp100) targeted T-cells when combined with topical SADBE. We will also combine fractional laser treatment plus anti-PD1 which, as we recently published, also triggers local recruitment of melanocyte-targeted T-cells. Aim 3 systematically uses our models to test varied dose/schedules of anti-inflammatory approaches that are commonly used in infants, to optimize SADBE’s nevus regression efficacy while minimizing cutaneous toxicity and inflammatory symptoms.

抽象的 先天性黑素细胞NEVI（CMN）可以达到巨大大小，转化为儿童黑色素瘤，从而触发 先发制人的手术引起了深远的发病率。在以前的资金期间，我们构建了四个CMN模型 测试旨在避免手术的诱导回归治疗方法。三个是基因工程的小鼠驱动的 由（Floxed）NRASQ61R ONCOGENE负责大多数人CMN，通过他莫昔芬 - 来自Tyr或DCT启动子的诱导或本构表达（各种强度）。第四模型是异种移植的 在免疫缺陷型小鼠上切除的人CMN。多种模型显示了长时间的黑色素瘤转化 潜伏期。诱导型NRASQ61R阐明了增殖的动力学与感觉奈夫相的动力学并进行了验证 在任一阶段取得成功的回归治疗方法。 tyr-cre; nrasq61r暴露了严重的人类中枢神经系统和皮肤 病变虽然较弱的DCT-CRE诱导了更典型​​的皮肤限制性巨型NEVI，其抑制剂的抑制剂， PI3K或CKIT诱导不完整（〜80％）的回归和组合完全消耗的视觉Nevocytes。局部 用促炎性触觉平方酸二丁酯（SADBE）诱导的完整视觉CMN处理 3剂后回归。该方案还完全防止了所有处理过的鼠类黑色素瘤转化 颈皮肤。我们发现局部SADBE需要巨噬细胞（不是B，T或NK细胞），因此可以在 人CMN异种移植物在SHO小鼠上（缺乏适应性免疫组织化学，但保留巨噬细胞）。 Sadbe 招募了鼠巨噬细胞到人异种移植物中，并将约90％的人类nevocytes递归。自从 SADBE已经在皮肤病学的其他地方使用，据报道诱导“脱位”，我们计划 临床测试。但是，由于在NIAMS的R01中不支持具有有效端点的临床试验，因此 我们正在寻求单独的资金来支持已经获得FDA/IND和IRB批准的临床试验。 虽然我们对在此资金期间发现的模型和策略感到兴奋，但我们认为这至关重要 以这一进展为基础，以增加实现这一毁灭性的真正治疗突破的可能性 儿童疾病。根据AIMS 1-3，使用我们的模型大胆寻求进一步提高效率并最小化 治疗的毒性：目标1深层和系统地发展了激酶抑制剂组合。它还测试MEK 抑制剂（MEKI）具有SADBE，因为最近暴露的个体效率和MEKI都被证明可以促进M1- 像（促炎）在M2样巨噬细胞分化上一样，一种机械特征，可能会增强 Sadbe的效率。 AIM 2寻求局部T细胞自适应免疫损害，以完成SADBE的 巨噬细胞诱导。我们已经显示了抗PD1触发黑素细胞（GP100）靶向T细胞的募集 当与局部sadbe结合使用时。我们还将结合分数激光治疗加抗PD1，这是我们 最近出版的内容也触发了当地招募靶向黑素细胞的T细胞。 AIM 3系统地使用我们的 测试在婴儿中常用的抗炎方法的各种剂量/时间表的模型 优化SADBE的Nevus回归效率，同时最大程度地减少皮肤毒性和炎症症状。