Project 1: Nanotherapies for Vemurafenib Resistant Melanoma

项目 1：维莫非尼耐药黑色素瘤的纳米疗法

• 批准号: 8960620
• 负责人: Leaf Huang
• 金额: $ 45.6万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8960620
• 关键词: Accounting; Address; Aftercare; Antigens; Basic Science; CCNE1 gene; Cancer Vaccines; Cells; Cessation of life; Cisplatin; Clinical Trials; Complementary DNA; Cutaneous Melanoma; Cytoplasm; DNA; Data; Dendritic Cells; Disease; Disease Resistance; Down-Regulation; Drug Formulations; Drug resistance; Early Diagnosis; Encapsulated; Endosomes; Exhibits; Gene Delivery; Genes; Glycolates; Goals; Growth; Human; Hyaluronic Acid; Immune; Immunotherapy; Life Expectancy; Lipid Bilayers; Lipids; MEKs; Malignant Neoplasms; Membrane Lipids; Messenger RNA; Metastatic Melanoma; Metformin; Modeling; Molecular Weight; Monophenol Monooxygenase; Mus; Mutate; Mutation; Nanotechnology; Neoplasm Metastasis; Nucleotides; PTEN gene; Particle Size; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Polyethyleneimine; Polymers; Prevention; Proteins; Resistance; Skin Cancer; Small Interfering RNA; Structure; Suppressor Genes; Therapeutic; Time; Toxic effect; Vaccine Therapy; Vaccines; Xenograft Model; abstracting; calcium phosphate; cancer therapy; clinically relevant; combination gene therapy; follow-up; gemcitabine; gene therapy; human FRAP1 protein; human SILV protein; in vivo; inhibitor/antagonist; innovation; lymph nodes; melanoma; melanoma-associated antigen; mouse model; mutant; nano; nanoparticle; nanotherapy; nanotool; neoplastic cell; novel; novel strategies; outcome forecast; polycation; raf Kinases; response; therapeutic vaccine; tool; tumor; tumor growth; vaccine development

Project 1 Abstract Cutaneous melanoma accounts for the majority (~75%) of skin cancer deaths. Despite the recent improvements in prevention and early detection, approximately 20% of patients with melanoma still die from the diseases. Patients with metastases have a poor prognosis and a very short life expectancy. About 50% of human melanoma harbor BRafV600 mutations. Vemurafenib (Vem), a specific inhibitor of the mutated BRaf, is very effective for treating metastatic melanoma. Unfortunately, drug resistance emerges 3-18 months post treatment due to alternative mechanisms that reactivate the MEK/ERK pathway. Vem resistant melanoma, however, exhibits elevated expression of melanoma associated antigens, gp100, MART-1 and Tyrosinase. Thus, the drug resistant disease provides opportunity for two independent, yet complementary, therapeutic approaches, i.e. synthetic lethality and cancer vaccine. To block the alternative Raf kinase, we have discovered that the suppressor gene Sprouty4, when delivered by LPH (lipid-polycation-hyaluronic acid) nanoparticles (NPs) developed in this group, could effectively down-regulate MEK/ERK pathway, leading to reduced growth rate in Vem resistant melanoma. We will follow up this novel gene therapy strategy by using a newly discovered lipopolyplex containing polymetformin (aim 1) to demonstrate the mechanism of the synthetic lethality (aim 2). Polymetformin is much less toxic than the equivalent polyethyleneimine and inhibits mTOR to further suppress the survival mechanism of the tumor cells. Since our Vem resistant tumor is also PTEN deficient, we will also develop a siRNA therapy to silence Akt. At the same time, we will use a BRaf mutant melanoma in a syngeneic mouse model to study tumor growth inhibition using a new mRNA vaccine formulation (aim 3). mRNAs of several melanoma associated antigens, including those up-regulated in Vem resistant tumor, will be formulated in LCP (lipid-calcium-phosphate) NPs and delivered to the dendritic cells in the lymph nodes. Preliminary data indicate that this approach was very effective in inducing an antigen-specific CTL response in the host and significantly inhibited the growth of the primary as well as the metastatic melanoma in the lymph nodes. We will finally combine the gene/drug therapy with the vaccine therapy to overcome Vem resistance in melanoma.

项目1 摘要 皮肤黑色素瘤占皮肤癌死亡的大部分（约 75%）。尽管 最近在预防和早期检测方面取得了进展，大约 20% 的患者患有 黑色素瘤仍然死于这些疾病。发生转移的患者预后较差， 预期寿命非常短。大约 50% 的人类黑色素瘤带有 BRafV600 突变。 维莫非尼 (Vem) 是一种突变 BRaf 的特异性抑制剂，对于治疗非常有效 转移性黑色素瘤。不幸的是，由于治疗后 3-18 个月，出现了耐药性 重新激活 MEK/ERK 通路的替代机制。 Vem 耐药性黑色素瘤， 然而，黑色素瘤相关抗原、gp100、MART-1 和 酪氨酸酶。因此，耐药性疾病为两种独立但尚未发生的疾病提供了机会。 补充性治疗方法，即合成致死剂和癌症疫苗。阻止 替代 Raf 激酶，我们发现抑制基因 Sprouty4，在传递时 通过该小组开发的LPH（脂质-聚阳离子-透明质酸）纳米颗粒（NP），可以 有效下调 MEK/ERK 通路，导致 Vem 耐药性生长速度降低 黑色素瘤。我们将通过使用新发现的一种新的基因治疗策略来跟进这种新的基因治疗策略 含有聚二甲双胍的脂多复合物（目标 1），以证明合成的机制 致死率（目标 2）。聚二甲双胍的毒性比同等的聚乙烯亚胺低得多， 抑制mTOR进一步抑制肿瘤细胞的生存机制。自从我们的维姆 耐药肿瘤也有PTEN缺陷，我们还将开发一种siRNA疗法来沉默Akt。在 同时，我们将在同基因小鼠模型中使用BRaf突变黑色素瘤来研究 使用新的 mRNA 疫苗配方抑制肿瘤生长（目标 3）。几个 mRNA 黑色素瘤相关抗原，包括那些在 Vem 抗性肿瘤中上调的抗原，将被 采用 LCP（脂质-钙-磷酸盐）纳米粒子配制并递送至树突状细胞 淋巴结。初步数据表明，这种方法对于诱导 宿主中抗原特异性 CTL 反应并显着抑制原代细胞的生长 以及淋巴结中的转移性黑色素瘤。我们最终将基因/药物结合起来 疫苗疗法可克服黑色素瘤中的 Vem 耐药性。