Aptamer mediated targeting of Fanconi Anemia oral cancer initiating cells

适体介导的范可尼贫血口腔癌起始细胞的靶向

• 批准号: 9060921
• 负责人: Ananth V Annapragada
• 金额: $ 38.38万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-25 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9060921
• 关键词: Aftercare; Antineoplastic Agents; Behavior; Binding; Cancer Patient; Cancer cell line; Cell Fraction; Cell Survival; Cells; Charge; Checkpoint kinase 1; Clinic; Clinical; Cytosol; Data; Development; Diagnosis; Disease-Free Survival; Distant; E-Selectin; Endosomes; Evolution; Exhibits; Fanconi' s Anemia; Gene Silencing; Gene Targeting; General Population; Genes; Goals; Head and neck structure; Hepatocyte; Hepatotoxicity; Hereditary Neoplastic Syndromes; In Vitro; Inherited; Kupffer Cells; Lead; Libraries; Ligands; Link; Lipids; Liposomes; Liver; Maintenance; Malignant Epithelial Cell; Malignant Neoplasms; Mediating; Methodology; Modality; Modeling; Mus; Oral; Patients; Pharmaceutical Preparations; Process; Property; Proteins; Public Health; Publishing; RNA Interference; Recurrence; Recurrent disease; Relapse; Risk; Role; Site; Small Interfering RNA; Specificity; Squamous cell carcinoma; Syndrome; System; TP53 gene; Techniques; Testing; Therapeutic; Therapeutic Agents; Time; Translating; Translations; Treatment Efficacy; Tumorigenicity; Xenograft procedure; aldehyde dehydrogenases; aptamer; base; cancer cell; cancer recurrence; cancer stem cell; cancer therapy; cell type; cellular engineering; chemoradiation; chemotherapy; gemcitabine; human stem cells; imaging agent; improved; in vivo; insight; keratinocyte; knock-down; malignant mouth neoplasm; mortality; mouth squamous cell carcinoma; nanoparticle; neoplastic cell; novel; outcome forecast; overexpression; public health relevance; response; screening; skills; targeted delivery; therapy outcome; therapy resistant; tumor; uptake

DESCRIPTION (provided by applicant): We seek to use a novel cell-internalization SELEX (Systemic Evolution of Ligands by Exponential enrichment) to isolate thioaptamer-conjugated liposomal nanoparticles (TA-NP) that internalize and deliver its siRNA into the cytosol of tumor initiating cells (TIC) of oral squamous cell carcinoma, minimizing off-target delivery. In vivo delivery and release of siRNA from the endosomes to cytosol remain the two biggest obstacles for translating anticancer siRNA drugs to the clinic. Oral squamous cell carcinoma commonly known as oral cancer (OC) is the most common malignancy of head and neck with high global public health impact. Fanconi anemia (FA) is a hereditary cancer syndrome that predisposes one to OC. Evidence points to TIC as the key driver of field cancerization, resistance to therapy and disease relapse. A therapeutic vehicle that selectively targets and delivers anticancer drugs to TIC offers great promise for OC treatment. Our data and published studies show: (1).TIC is more enriched in FA-OC than sporadic OC; (2) Chk1 and CD147 promote TIC survival and chemoresistance and its overexpression contribute to poor prognosis in OC. The main goal of this application is to use FA- OC-TIC as target cells to develop TA- NP for the cytosolic delivery of siRNA. We hypothesize that isolated TA-NP will facilitate targeted delivery of siRNA to OC-TIC cytosol, and silence CD-147/Chk1 in OC-TIC's, disrupting their niche and rendering them chemosensitive and susceptible to elimination by chemotherapy with gemcitabine. Aims:(1): Elucidate the effect of RNAi mediated silencing of CD147/Chk1 in FA-OC-TIC; (2A): Select a list of TA-Liposomal NP (TA-NP) specific for TIC fraction of FA-OC cells, while avoiding hepatocyte uptake, using a positive and negative cell-uptake based SELEX; (2B): Determine the selected TA-NP's targeting specificity and therapeutic efficacy in in vitro and in murine OC xenografts. We will isolate ALDH+ TIC in FA-OC cell lines and examine the effects of siRNA-mediated silencing of CD147 and Chk1. (2) We will use a modified conjugate SELEX with positive and negative selections to identify OC-TIC-specific internalizing TA-NP. (3) We will validate TIC-targeting specificity and silencing efficacy of isolated TA-NP-sRNA in vitro and in murine orthotopic FA-OC xenografts. RNAi mediated inhibition of CD147 and Chk1 in OC-TIC will yield valuable insight into their potential role in the tumor propagating and chemoresistant properties of OC-TIC. The proposed TA-NP will have important clinical potentials for in vivo delivery of therapeutic and imaging agents targeting the OC-TIC.

描述（由申请人提供）：我们寻求使用新的细胞内化SELEX（通过指数富集的配体系统进化）来分离硫代适体缀合的脂质体纳米颗粒（TA-NP），其内化并将其siRNA递送到口腔鳞状细胞癌的肿瘤起始细胞（TIC）的胞质溶胶中，最小化脱靶递送。siRNA从内体到胞质溶胶的体内递送和释放仍然是将抗癌siRNA药物转化为临床的两个最大障碍。口腔鳞状细胞癌（Oral squamous cell carcinoma，OC）是头颈部最常见的恶性肿瘤，对全球公共卫生影响很大。范可尼贫血（FA）是一种遗传性癌症综合征，易患OC。有证据表明，TIC是实地癌变、治疗抵抗和疾病复发的关键驱动因素。选择性靶向TIC并将抗癌药物递送至TIC的治疗载体为OC治疗提供了巨大的希望。我们的数据和已发表的研究表明：（1）TIC在FA-OC中比散发性OC更富集;（2）Chk 1和CD 147促进TIC的生存和化疗耐药性，其过表达有助于OC的不良预后。本申请的主要目标是使用FA-OC-TIC作为靶细胞来开发用于siRNA的胞质递送的TA-NP。我们假设，分离的TA-NP将促进siRNA靶向递送至OC-TIC胞质溶胶，并沉默OC-TIC中的CD-147/Chk 1，破坏它们的生态位，并使它们对吉西他滨化疗敏感并易于消除。目的：（1）：阐明FA-OC-TIC中RNAi介导的CD 147/Chk 1沉默的作用;（2A）：使用基于阳性和阴性细胞摄取的SELEX，选择对FA-OC细胞的TIC级分特异性的TA-脂质体NP（TA-NP）的列表，同时避免肝细胞摄取;（2B）：确定所选TA-NP在体外和鼠OC异种移植物中的靶向特异性和治疗功效。我们将在FA-OC细胞系中分离ALDH+ TIC，并检查siRNA介导的CD 147和Chk 1沉默的效果。(2)我们将使用具有阳性和阴性选择的修饰的缀合物SELEX来鉴定OC-TIC特异性内化TA-NP。(3)我们将在体外和鼠原位FA-OC异种移植物中验证分离的TA-NP-sRNA的TIC靶向特异性和沉默功效。RNAi介导的OC-TIC中CD 147和Chk 1的抑制将产生对它们在OC-TIC的肿瘤增殖和化学抗性特性中的潜在作用的有价值的洞察。提出的TA-NP将具有重要的临床潜力，在体内传递的治疗和成像剂的目标OC-TIC。