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.

摘要 先天性黑色素细胞痣(CMN)可以达到巨大的尺寸，转化为儿童黑色素瘤，从而引发 先发制人的手术导致了严重的并发症。在之前的资助期，我们构建了四个CMN模型 测试旨在避免手术的回归诱导治疗。其中三只是基因工程小鼠 通过(漂白的)NRASQ61R癌基因，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启动子的可诱导或组成性表达(强度不同)。第四种模型是异种移植。 切除免疫缺陷小鼠的人巨噬细胞集落刺激因子。几个模型显示长时间后黑色素瘤转化 延迟。可诱导的NRASQ61R阐明了增殖期与衰老期雀斑的动力学，并验证了 倒退治疗在任何一个阶段都能取得成功。TYR-CRE；NRASQ61R表现出严重的类人中枢神经系统和皮肤 皮损，而较弱的DCT-CRE诱导了更典型的皮肤限制性巨痣，MEK的抑制剂， PI3K或cKit诱导的不完全退化(~80%)和联合完全耗尽的视觉新生细胞。热门话题 促炎性半抗原方酸二丁酯治疗完全性视觉CMN 3次给药后消退。在所有治疗的范围内，该方案还完全防止了小鼠黑色素瘤的转化 雀斑皮肤。我们发现，外用SADBE需要巨噬细胞(不是B、T或NK细胞)，所以它可以在 将人CMN异种移植到缺乏获得性免疫但保留巨噬细胞的SHO小鼠身上。SADBE 将小鼠巨噬细胞招募到人的异种移植中，使约90%的人新生细胞退化。自.以来 SADBE已经在皮肤科的其他地方使用过，据报道会导致“色素脱失”，我们计划 对其进行临床测试。然而，由于NIAMS的R01中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经获得FDA/IND和IRB批准的临床试验。 虽然我们对在这一资助期间发现的模型和战略感到兴奋，但我们认为这是至关重要的 在这一进展的基础上，增加实现真正的治疗突破的可能性 儿童疾病。相应地，目标1-3利用我们的模型大胆寻求进一步的增强功效和最小化 治疗的毒性：目的1深入和系统地开发激酶抑制剂组合。它还测试MEK 抑制剂(Meki)与SADBE一起使用，因为两者都显示出个体疗效，Meki最近被证明能促进M1- 像(促炎)超过M2样巨噬细胞分化，这是一种可能增强 SADBE的功效。AIM 2寻求局部T细胞适应性免疫招募，以补充SADBE 巨噬细胞诱导。我们已经证明了抗PD1可以触发针对黑素细胞(Gp100)的T细胞的募集 当与外用SADBE联合使用时。我们还将结合部分激光治疗和抗PD1，因为我们 最近发表的，也触发了黑素细胞靶向T细胞的局部招募。AIM 3系统地使用我们的 测试婴儿常用的不同剂量/时间表的抗炎方法的模型，以 优化SADBE的雀斑消退功效，同时将皮肤毒性和炎症症状降至最低。