Therapeutic strategies for treatment of giant congenital melanocytic nevi

巨大先天性黑素细胞痣的治疗策略

基本信息

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
项目类别：
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

项目摘要

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.

抽象的先天性黑色素细胞痣 (CMN) 可以达到巨大尺寸，转变为儿童黑色素瘤，从而引发在之前的资助期间，我们构建了四个 CMN 模型。测试旨在避免手术的回归诱导疗法，其中三种是基因工程小鼠。由（floxed）NRASQ61R癌基因负责，负责大多数人类CMN，通过他莫昔芬靶向黑素细胞- 来自 Tyr 或 Dct 启动子的诱导型或组成型表达（不同强度）第四种模型是异种移植。在免疫缺陷小鼠身上切除人类 CMN，一些模型在长时间后表现出黑色素瘤转化。诱导型 NRASQ61R 阐明了增殖痣阶段与衰老痣阶段的动力学并得到验证。 Tyr-Cre 的回归治疗取得成功；NRASQ61R 显示出严重的类人中枢神经系统和皮肤。病变，而较弱的 DCT-Cre 诱发更典型的皮肤限制性巨大痣，MEK 抑制剂， PI3K 或 cKIT 诱导不完全 (~80%) 消退，并且组合完全耗尽视觉神经细胞。使用促炎半抗原方酸二丁酯 (SADBE) 治疗可诱导完全视觉 CMN 在所有治疗中，该方案还完全阻止了小鼠黑色素瘤的转化。我们发现局部 SADBE 需要巨噬细胞（不是 B、T 或 NK 细胞），因此可以在皮肤上进行测试。人类 CMN 异种移植到 SHO 小鼠（缺乏适应性免疫但保留巨噬细胞）。将小鼠巨噬细胞招募到人类异种移植物中，并消退了约 90% 的人类神经细胞。 SADBE 已在皮肤科的其他地方使用，据报道会引起“色素脱失”，我们计划然而，由于 NIAMS 的 R01 不支持具有疗效终点的临床试验，我们正在寻求单独的资金来支持这项已获得 FDA/IND 和 IRB 批准的临床试验。虽然我们对本次融资期间的模型和未发现的策略感到兴奋，但我们认为这是至关重要的在此基础上再接再厉，增加针对这种毁灭性的疾病实现真正治疗突破的可能性因此，目标 1-3 使用我们的模型大胆寻求进一步提高疗效并最小化。治疗的毒性：目标1深入、系统地开发激酶抑制剂组合，它还测试MEK。抑制剂 (MEKi) 与 SADBE，因为两者都表现出各自的功效，并且 MEKi 最近被证明可以促进 M1- 类似于 M2 样巨噬细胞分化（促炎症），这一机制特征可能会增强 SADBE 的目标 2 寻求局部 T 细胞免疫适应性招募，以补充 SADBE 的功效。我们已经证明抗 PD1 会触发黑色素细胞 (gp100) 靶向 T 细胞的募集。当与局部 SADBE 结合时，我们还将结合点阵激光治疗和抗 PD1。最近发表的 Aim 3 也系统地使用了我们的方法，触发了针对黑色素细胞的 T 细胞的局部招募。模型来测试婴儿常用的不同剂量/时间表的抗炎方法，优化 SADBE 的痣消退功效，同时最大限度地减少皮肤毒性和炎症症状。