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 皮肤黑色素瘤占皮肤癌死亡的大多数（约75%）。尽管 最近在预防和早期发现方面的改进，大约20%的患者 黑色素瘤仍然死于这些疾病。有转移的患者预后差， 非常短的预期寿命。约50%的人黑素瘤具有BRafV 600突变。 Vemurafenib（Vem）是突变BRaf的特异性抑制剂， 转移性黑素瘤不幸的是，耐药性在治疗后3-18个月出现， 重新激活MEK/ERK通路的替代机制。Vem抗性黑素瘤， 然而，其表现出黑素瘤相关抗原gp 100、MART-1和 酪氨酸酶。因此，耐药性疾病提供了两个独立的，但 互补的治疗方法，即合成致死性和癌症疫苗。阻止 作为Raf激酶的替代物，我们发现抑制基因Sputy 4， 通过LPH（脂质-聚阳离子-透明质酸）纳米颗粒（NPs）， 有效地下调MEK/ERK通路，导致Vem抗性中生长速率降低 黑素瘤我们将通过使用新发现的 含有聚二甲双胍的脂质多聚物（AIM 1），以证明合成的机制 致命性（目标2）。聚二甲双胍的毒性远低于等效的聚乙烯亚胺， 抑制mTOR以进一步抑制肿瘤细胞的存活机制。自从我们的Vem 由于耐药肿瘤也是PTEN缺陷的，我们还将开发siRNA疗法来沉默Akt。在 同时，我们将使用BRaf突变黑色素瘤在同基因小鼠模型中进行研究 使用新的mRNA疫苗制剂抑制肿瘤生长（目的3）。几种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 在LCP（脂质-磷酸钙）纳米颗粒中配制并递送到树突细胞中， 淋巴结初步数据表明，这种方法在诱导 抗原特异性CTL反应，并显着抑制原代细胞的生长， 以及淋巴结转移性黑色素瘤我们最终将联合收割机 用疫苗疗法来克服黑色素瘤中的Vem抗性。