A Novel Targeted Nanomedicine Delivering MicroRNA-30-5p Replacement Therapy for Multi-drug Resistant Cancer Treatment

摘要： 在这项SBIR应用中，miRecule建议开发一种基于microRNA的miR-30-5p的治疗模拟物 (miRecule候选MC-30)用于治疗多药耐药(MDR)癌症。头颈部鳞状细胞 细胞癌(HNSCC)是第六大常见癌症。超过一半的患者患有晚期 疾病III期或IV期，平均5年生存率~40%。HNSCC肿瘤具有高水平的基因 突变会导致高度的肿瘤异质性和耐药性。Mir-30-5p的表达被广泛抑制 在肿瘤组织中，约30%的HNSCC存在MIR30基因缺失。基于MIR的疗法提供了一种 靶向主要致癌途径和潜在的多药耐药癌症治疗 抑制内在或获得性抗性的机制。EGFR靶向治疗经常被 代偿性生长因子受体(GFRs)MET和IGF1R过表达。我们已经把这部小说写成了 发现miR-30-5P可以靶向并抑制所有这三种GFR。我们设计的基本原理是 由于MIR-30-5P MIMIC具有治疗异质性晚期HNSCC的能力，因此它将在治疗异质性晚期HNSCC方面具有优势 不仅调节EGFR，还调节MET、IGF-1R和其他二十多个证实被解除调节的mRNAs 在肿瘤组织中，与过度增殖、黏附、迁移、细胞外基质重塑和 差异化。我们早期的研究表明，经过化学修饰的miR-30-5P的模拟物，表现出显著的 与生物microRNA相比，提高了疗效，在临床前有效地抑制了体内肿瘤生长 HNSCC模型。 核酸疗法的一个重大障碍是缺乏一种有效的方法来具体地传递它们 靶向癌细胞，尤其是在转移阶段。为了克服这一障碍，我们聘请了两个独特的 战略。首先，我们的miR-30-5P的化学修饰模拟物极大地提高了核酸酶的稳定性和 活性，减少细胞中产生所需效果所需的模拟物的量，以及 延长响应持续时间。其次，我们的模拟物被包裹在一个经过临床验证的肿瘤中- 靶向脂质体纳米给药系统(SCL)。该纳米复合体携带抗单链抗体片段。 转铁蛋白受体(TFR)，它在大多数癌细胞表面上调，并通过 受体介导的内吞作用。在这项第一阶段的SBIR研究中，我们的目标是描述我们的 HNSCC细胞中铅核酸酶稳定的模拟物、多个致癌基因的靶向和信号转导 并测试我们的假设，即它可以在体外更有效地对抗耐药性(目标1)。我们会 在西妥昔单抗和顺铂耐药的HNSCC原位移植小鼠模型中进行模拟试验 直接与顺铂和西妥昔单抗的护理标准进行比较。我们还将评估潜在的毒性和有害物质- 目标效果。(AIM2)这些研究的成功完成将证明MC-30作为一种 未来PK、TOX、ADME和其他动物研究中需要使用的候选药物和给药方案 对于IND套餐。一旦FDA批准并上市，MC-30将提供有效的治疗 MDR、HNSCC，或许还有其他目前尚无治疗选择的癌症。

Abstract: In this SBIR application miRecule proposes to develop a microRNA-based therapeutic mimic of miR-30-5p (miRecule candidate MC-30) for the treatment of multi-drug resistant (MDR) cancers. Head and Neck Squamous Cell Carcinoma (HNSCC) is the 6th most common form of cancer. Greater than half of patients present with late stage III or IV disease, with an average 5-year survival rate of ~40%. HNSCC tumors have high levels of genetic mutations leading to high tumor heterogeneity and drug resistance. miR-30-5p expression is widely repressed in tumor tissues, and MIR30 gene deletion is observed in ~30% of HNSCCs. miR-based therapeutics offer a disruptive MDR cancer treatment by targeting both the primary oncogenic pathways and potentially suppressing mechanisms of intrinsic or acquired resistance. EGFR targeted therapy is often resisted by overexpression of the compensating growth factor receptors (GFRs) MET and IGF1R. We have made the novel discovery that miR-30-5p can target and repress all three of these GFRs. The rationale for our design is that a miR-30-5p mimic will be superior in its ability to treat heterogeneous late-stage HNSCC due to its ability to regulate not only EGFR, but also MET, IGF-1R, and over two dozen other mRNAs confirmed to be deregulated in tumor tissue and associated with over-proliferation, adhesion, migration, extracellular matrix remodeling, and differentiation. Our early study shows that a chemically modified mimic of miR-30-5p, exhibiting significantly improved efficacy over the biological microRNA, was effective at inhibiting tumor growth in vivo in a preclinical model of HNSCC. A significant hurdle for nucleic acid therapeutics is a lack of an efficient means of delivering them specifically to target cancer cells, especially at the metastatic stage. To overcome this obstacle, we employ two unique strategies. First, our chemically modified mimic of miR-30-5p has vastly improved nuclease stability and activity, decreasing the amount of mimic that needs to be present in the cell to produce the desired effect, and extending the duration of response. Second, our mimics are encapsulated in a clinically validated tumor- targeted liposomal nanodelivery system (scL). The nanocomplex carries an scFv antibody fragment against the transferrin receptor (TfR), which is up-regulated on the surface of most cancer cells and triggers uptake via receptor-mediated endocytosis. In this Phase I SBIR study we aim to we will characterize the half-life of our lead nuclease-stabilized mimic in HNSCC cells, targeting of multiple oncogenic mRNAs and signaling pathways, and test our hypothesis that it can more effectively combat drug resistance in vitro (Aim 1). We will test this mimic in a cetuximab and cisplatin-resistant orthotopic xenograft mouse model of HNSCC and compare directly with standard of care cisplatin and cetuximab. We will also assess potential toxicity and off- target effects. (AIM2) Successful completion of these studies will demonstrate the feasibility of MC-30 as a drug candidate and dosing regimen to be used in future PK, TOX, ADME, and other animal studies as required for an IND package. Once FDA approved and commercially available MC-30 will offer an effective treatment of MDR HNSCC and perhaps other cancers for which there are currently no treatment options.