Molecularly Targeted Probes for Photodynamic Therapy and Imaging of Breast Cancer

用于乳腺癌光动力治疗和成像的分子靶向探针

• 批准号: 10358832
• 负责人: Hans F. Schmitthenner
• 金额: $ 45.77万
• 依托单位: ROCHESTER INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10358832
• 关键词: Affinity; Aftercare; Apoptosis; Area; Binding; Biodistribution; Biological Assay; Biological Markers; Biology; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Patient; Breast Cancer Treatment; Breast Cancer cell line; Breast Cancer therapy; Cell Death; Cell Line; Cell Survival; Cells; Cellular Assay; Cellular biology; Chemistry; Clinical Treatment; Collaborations; Confocal Microscopy; Coupling; Dimensions; Disease; Disease Progression; Dyes; Endocytosis; Ensure; Epidemic; Evaluation; Fatty acid glycerol esters; Female; Fluorescence; Fluorescence Microscopy; Goals; Hormones; Image; In Vitro; Injections; Keratin; Lasers; MCF10A cells; MDA MB 231; Measurement; Measures; Methodology; Methods; Modality; Molecular Target; Monitor; Mus; Near-infrared optical imaging; Operative Surgical Procedures; Optics; Outcome; PUVA Photochemotherapy; Patients; Peptide Synthesis; Peptides; Phase; Photosensitizing Agents; Property; Protocols documentation; Reporting; Residual state; Site; Solid; Specificity; Students; Testing; Time; Tissues; Treatment outcome; Tumor Volume; Universities; anticancer research; base; breast imaging; breast lumpectomy; cancer imaging; cancer therapy; cyanine dye 5; design; educational atmosphere; efficacy evaluation; efficacy testing; experimental study; fluorescence imaging; fluorescence-guided surgery; imaging agent; in vivo; in vivo evaluation; in vivo monitoring; malignant breast neoplasm; mammary; minimally invasive; molecular imaging; near infrared dye; neoplastic cell; novel; operation; overexpression; receptor; receptor binding; response; side effect; targeted agent; treatment optimization; triple-negative invasive breast carcinoma; tumor; undergraduate student

Summary: Molecularly Targeted Probes for Photodynamic Therapy and Imaging of Breast Cancer The goal of this proposal is to test the hypothesis that small, peptide-based molecularly targeted probes containing a near infrared (NIR) dye and a photosensitizer (PS) dye will be effective for fluorescence imaging and photodynamic therapy (PDT) of breast cancer (BrCa). This novel treatment method will give patients access to an unprecedented therapy option, without the burden of side effects from traditional BrCa therapy. In particular, the application to fluorescence-guided lumpectomy combined with PDT in the same operation is envisioned as a remedy for the current epidemic of re-operation as it will provide a means of effectively treating residual BrCa cells in the margins. The strategy relies on a targeting deca-peptide, 18-4, reported to be an effective targeting agent for the keratin receptor, KRT1, a biomarker that is independent of hormone expression in BrCa cells. Students in our labs have developed a synthesis of 18-4 by a solid phase peptide synthesis (SPPS) and have merged that with a modular method we developed for the synthesis of BrCa-targeted molecular imaging agents (TMIAs). This method will provide analogous molecularly targeted photosensitizer (MTPS) probes for PDT and dual TMIA-MTPS probes for guiding surgery and PDT. Students at RIT will use confocal fluorescence microscopy (CFM) to evaluate the affinity of the 18-4-based TMIAs and dual TMIA-MTPS probes to two triple negative BrCa cell lines, MDA-MB-231 and EMT6, utilizing non-cancerous MCF10A as a control. The in vitro efficacy of PDT in the same cell lines will next be investigated using a single MTPS probe. To ensure that single MTPS and dual TMIA-MTPS probes are equally effective, PDT will be investigated in vitro in the MDA-MB-231 and EMT6 cells lines, using a laser supplied by our collaborator at the U of R. The mechanism of cell death will be studied using CFM and apoptosis assays. The strategy includes testing the binding affinity of the single and dual probes to EMT6 tumors in mice by in vivo NIR imaging, then testing efficacy in PDT experiments after systemic injection of the targeted dual TMIA-MTPS probe. Therapy will be monitored by tumor dimensions and stereo-fluorescence microscopy to verify efficacy of targeted and untargeted probes, with and without PDT. The combined in vitro and in vivo experiments will provide an effective strategy to develop single and dual probes for the PDT of BrCa. A key aspect of this proposal is the participation of three undergraduates from RIT in each year, one of whom will participate in the in vivo testing at the U of R, one in synthesis and one in the in vitro analysis at RIT. The collaboration highlights expertise in synthesis, cell biology, and confocal microscopy at RIT, and in cancer imaging and photodynamic therapy at the U of R. These combine to provide a vibrant learning environment in cancer research for undergraduates, with the achievable goal of revolutionizing treatment of BrCa by developing small, peptide-based TMIA, MTPS, and dual TMIA-MTPS probes to transform BrCa therapy by the use of molecularly targeted fluorescence-guided PDT and provide a breakthrough in the treatment and cure for BrCa patients.

综述：用于乳腺癌光动力治疗和成像的分子靶向探针 这项提议的目的是检验一种假设，即小的、基于肽的分子靶向探针 含有近红外（NIR）染料和光敏剂（PS）染料的荧光成像将有效 和乳腺癌（BrCa）的光动力疗法（PDT）。这种新的治疗方法将使患者 获得前所未有的治疗选择，而没有传统BrCa治疗的副作用负担。在 特别地，在同一手术中应用荧光引导的肿块切除术结合PDT， 设想作为一种补救措施，目前的流行病再次手术，因为它将提供一种手段，有效地治疗 边缘残留BrCa细胞该策略依赖于靶向十肽，18-4，据报道是一种 角蛋白受体KRT 1的有效靶向剂，KRT 1是一种不依赖于激素表达的生物标志物 在BrCa细胞中。我们实验室的学生通过固相肽合成（SPPS）开发了18-4的合成方法 并将其与我们开发的用于合成BrCa靶向分子的模块化方法相结合， 显像剂（TMIA）。这种方法将提供类似的分子靶向光敏剂（MTPS） PDT探头和用于引导手术和PDT的双TMIA-MTPS探头。RIT的学生将使用共焦 荧光显微镜（CFM）以评价基于18-4的TMIA和双TMIA-MTPS探针的亲和力 两种三阴性BrCa细胞系MDA-MB-231和EMT 6，使用非癌性MCF 10A作为对照。 接下来将使用单一MTPS探针研究PDT在相同细胞系中的体外功效。到 为了确保单MTPS和双TMIA-MTPS探针同样有效，将在体外研究PDT， MDA-MB-231和EMT 6细胞系，使用我们的合作者在R. U.的 将使用CFM和凋亡测定来研究细胞死亡的机制。该战略包括测试 通过体内近红外成像，然后测试单探针和双探针与小鼠EMT 6肿瘤的结合亲和力 在PDT实验中，在全身注射靶向双TMIA-MTPS探针后，治疗将 通过肿瘤尺寸和立体荧光显微镜监测，以验证靶向和 非靶向探针，有和没有PDT。体外和体内实验的结合将提供一个 有效的策略，开发单探针和双探针的PDT的BrCa。该提案的一个关键方面是， 每年RIT的三名本科生参与，其中一人将参与体内测试 在R的U，一个在合成，一个在RIT的体外分析。该合作突出了以下方面的专业知识 合成，细胞生物学和共聚焦显微镜，以及癌症成像和光动力治疗， R的U。这些联合收割机为本科生提供了一个充满活力的癌症研究学习环境， 随着通过开发小的基于肽的TMIA来革新BrCa治疗的可实现目标， MTPS和双TMIA-MTPS探针，通过使用分子靶向 荧光引导的PDT，为BrCa患者的治疗和治愈提供了突破。