Molecular Targeted Photoimmunotherapy for Bladder Cancer

膀胱癌的分子靶向光免疫治疗

• 批准号: 10014685
• 负责人: Piyush Agarwal
• 金额: $ 28.89万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10014685
• 关键词: Accounting; Address; Animals; Antibodies; Antibody-drug conjugates; Apoptosis; Attention; BCG Live; Bladder; Bladder Neoplasm; Cancer Model; Cancer cell line; Catheters; Cause of Death; Cell Death; Cell Line; Cessation of life; Clinical; Colon Carcinoma; Contracture; Cost of Illness; Cutaneous; Cystoscopes; Death Rate; Diagnosis; Disease; ERBB2 gene; Epidermal Growth Factor Receptor; Equipment; FGFR3 gene; Fiber; Future; Goals; Growth Factor Receptors; Human; Immune system; Immunocompetent; Immunofluorescence Immunologic; Immunotherapy; In Vitro; Individual; Infrared Rays; Lasers; Light; Low Prevalence; Luciferases; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Manuscripts; Methods; Mitomycins; Modeling; Molecular; Molecular Target; Monoclonal Antibodies; Mus; Muscle; Mutation; Mutation Analysis; Necrosis; Operative Surgical Procedures; PD-1/PD-L1; PIK3CA gene; PUVA Photochemotherapy; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phase I Clinical Trials; Photosensitization; Positive Lymph Node; Pre-Clinical Model; Prevalence; Property; Publications; Publishing; Recurrence; Reporting; Research; Role; Signal Pathway; Signal Transduction; Surface; Technology; Testing; Therapeutic; Transurethral Resection; Tyrosine Kinase Inhibitor; United States; Urologic Oncology; Viral; Woman; Work; antibody conjugate; cancer cell; carcinogenesis; cytotoxicity; flexibility; humanized monoclonal antibodies; immunogenic cell death; implantation; improved; in vivo; interest; men; molecular imaging; molecular targeted therapies; muscle invasive bladder cancer; non-muscle invasive bladder cancer; novel; novel strategies; overexpression; particle; photoimmunotherapy; pre-clinical; prevent; receptor; receptor expression; side effect; standard of care; success; targeted agent; targeted treatment; therapy development; tumor; tumor heterogeneity; urinary; urologic

Non-muscle-invasive bladder cancer has few available treatments. The mainstay of treatment is surgery with a transurethral resection of the bladder tumor but the recurrence rate is as high as 75%. Mitomycin C is given peri-operatively to reduce the recurrence rate and when disease is high grade, Bacillus Calmette-Guerin (BCG) is used to prevent both recurrence and progression. However, these treatments often fail and when successful, they work through a poorly understood mechanism. Therefore, attention has been directed towards molecular alterations in bladder cancer to identify novel targets. Mutation analysis of 145 urothelial tumors found that FGFR3 and PIK3CA mutations were most commonly found in low grade non-muscle-invasive tumor (Sjodahl et al, 2011). In fact, the FGFR3 mutation is found in up to 80% in low-grade bladder tumors and up to 50% of invasive tumors. In addition, over-expression or amplification of EGFR is seen in the majority of urothelial tumors (Rotterud et al, 2005). However, targeted therapy with the monoclonal antibodies has been disappointing in bladder cancer. Multiple studies have been conducted with EGFR-targeted monoclonal antibodies and tyrosine kinase inhibitors without any significant clinical benefit despite encouraging pre-clinical results (Black et al, 2012). Recently, an FGFR3 monoclonal antibody was effective but primarily in invasive bladder cancer cell lines instead of the low grade bladder cancer where mutations in FGFR3 are more commonly found (Qing et al, 2009). Despite having these well-defined targets in non-muscle-invasive bladder cancer, there has been little success with targeted therapy alone. Recently, colleagues from the molecular imaging branch demonstrated successfully that humanized monoclonal antibodies could be used to target infrared light activated compounds selectively into colon cancer cells. Upon introduction of infrared light, these compounds become activated to induce cell death (Mitsunaga et al, 2011). These antibody-infrared light activated drug conjugates are able to capitalize on the targetable property of the antibodies but rely on the cytotoxicity of the drug conjugated to the antibody and not the antibody itself. Given the prevalence of EGFR and FGFR3 mutations in non-muscle-invasive bladder cancer and the disappointing results with standard targeted therapy, we postulate that antibody-infrared light activated drug conjugates will have significant activity in inducing cell death selectively in bladder cancer cells. This is novel as an infrared light could be attached to existing urologic equipment such as a urinary catheter or flexible cystoscope and be introduced into the bladder to activate such conjugates instilled into the bladder should pre-clinical work suggest a benefit of these agents in bladder cancer. We will further investigate molecular targeted photoimmunotherapy in bladder cancer by specifically targeting EGFR, FGFR3, and other related targets. The objectives are: 1) To profile bladder cancer cell lines by surface receptor expression and mutations so as to better understand their role as models of urothelial cancer; 2) to establish in vitro efficacy of photodynamic therapy (specifically photoimmunotherapy); 3) to determine the mechanism of cell death caused by photodynamic therapy (PIT); 4) to use antibody-drug conjugates in animals and humans alone and in synergistic combinations; 5) evaluate for immunogenic cell death. Our key accomplishments are as follows: 1) we have obtained a large number of bladder cancer cell lines and we have personally developed three cell lines within the Urologic Oncology Branch from patient tumors. These cell lines have been thoroughly characterized using a mutational array. Furthermore, several cell lines have been successfully transfected with luciferase and we have begun orthotopic implantation with these lines; 2) we have thoroughly profiled several bladder cancer cell lines for surface expression of growth factor receptors for which monoclonal antibodies are available. We are now embarking on characterizing them for FGFR3 expression; 3) we have confirmed the efficacy of photoimmunotherapy (aim 2) and determined that necrosis is the likely mechanism of action (aim 3). We have completed animal studies (aim 4) and confirmed presence of EGFR and other targets in actual human tumors by quantitative immunofluorescence. We published our first manuscript describing our in vitro and in vivo work in Molecular Cancer Therapeutics. We just completed combination antibody-drug conjugates using EGFR and HER-2 targeted approach concurrently (aim 4) and are submitting the results for publication. In this upcoming report, we have shown that combination of two antibody-drug conjugates can be helpful in tumors with lower expression of each individual target. We also have a review article on photodynamic therapy and photoimmunotherapy in press. We are now trying to establish that photoimmunotherapy results in immunogenic cell death which can prime the immune system (aim 5). After confirming our findings in an immunocompetent mouse, we will combine photoimmunotherapy (single antibody-drug conjugate and multiple antibody-drug conjugates) with immunotherapy (PD-1/PD-L1 therapy - aim; 4). Finally, we hope to partner with the NCI or a commercial collaborator to introduce such therapy to patients in a phase I clinical trial.

非肌肉侵入性膀胱癌几乎没有可用的治疗方法。治疗的主要手术是膀胱肿瘤的尿道切除术，但复发率高达75％。丝霉素C进行围手术期以降低复发率，而当疾病是高级时，Calmetles Calmette-Guerin（BCG）用于防止复发和进展。但是，这些治疗方法常常失败，并且在成功的情况下，它们是通过知之甚少的机制来工作的。因此，注意力集中在膀胱癌中的分子改变，以鉴定新的靶标。对145种尿路上皮肿瘤的突变分析发现，在低级非肌肉侵入性肿瘤中，FGFR3和PIK3CA突变最常发现（Sjodahl等，2011）。实际上，在低级膀胱肿瘤中发现了多达80％的FGFR3突变，高达50％的浸润性肿瘤。另外，在大多数尿路上皮肿瘤中都可以看到EGFR的过表达或扩增（Rotterud等，2005）。但是，用单克隆抗体的靶向治疗对膀胱癌令人失望。尽管鼓励了临床前结果，但已使用EGFR靶向的单克隆抗体和酪氨酸激酶抑制剂进行了多项研究（Black等，2012）。最近，FGFR3单克隆抗体是有效的，但主要在浸润性膀胱癌细胞系中而不是低级膀胱癌中，其中FGFR3中的突变更常见（Qing等，2009）。尽管在非肌肉侵入性膀胱癌中具有这些定义明确的靶标，但仅针对靶向疗法就没有成功。最近，来自分子成像分支的同事成功证明了人源化的单克隆抗体可用于选择性地靶向红外活性化合物，以选择性地靶向结肠癌细胞。引入红外光后，这些化合物被激活以诱导细胞死亡（Mitsunaga等，2011）。这些抗体 - 含光活化的药物缀合物能够利用抗体的目标特性，但依赖于与抗体共轭的药物的细胞毒性，而不是抗体本身。鉴于非肌肉侵入性膀胱癌中EGFR和FGFR3突变的患病率以及标准靶向疗法的令人失望的结果，我们假设抗体含量的光活化的药物共轭物在选择性地诱导细胞死亡的膀胱癌细胞中具有显着的活性。这是新颖的，因为红外光可以连接到现有的泌尿外科设备，例如尿道导管或柔性膀胱镜，并被引入膀胱中，以激活灌输到膀胱的这种结合物，应表明这些药物在膀胱癌中有好处。我们将通过专门针对EGFR，FGFR3和其他相关靶标，进一步研究膀胱癌中的分子靶向摄影疗法。目的是：1）通过表面受体表达和突变来介绍膀胱癌细胞系，以更好地理解它们作为尿路上皮癌模型的作用； 2）建立光动力疗法的体外功效（特别是摄影疗法）； 3）确定光动力疗法引起的细胞死亡机制（PIT）； 4）单独使用动物和人类和协同组合中使用抗体 - 药物缀合物； 5）评估免疫原性死亡。我们的主要成就如下：1）我们获得了大量的膀胱癌细胞系，并且我们在患者肿瘤的泌尿外科肿瘤分支中亲自开发了三个细胞系。这些细胞系已通过突变阵列彻底表征。此外，几种细胞系已成功地用荧光素酶转染，我们已经开始使用这些系的原位植入。 2）我们已经彻底介绍了几种膀胱癌细胞系，用于可用的单克隆抗体的生长因子受体的表面表达。现在，我们正在开始表征它们以表达FGFR3。 3）我们已经证实了摄影疗法的功效（AIM 2），并确定坏死是可能的作用机理（AIM 3）。我们已经完成了动物研究（AIM 4），并通过定量免疫荧光确认了实际人类肿瘤中EGFR和其他靶标的存在。我们发表了第一个描述我们在分子癌治疗中的体外和体内工作的手稿。我们刚刚使用EGFR和HER-2靶向方法（AIM 4）完成了组合抗体 - 药物缀合物，并正在提交结果出版。在这份即将发布的报告中，我们已经表明，两种抗体 - 药物结合物的组合可能有助于每个靶标表达较低的肿瘤。我们还收到了一篇有关光动力疗法和摄影疗法的评论文章。现在，我们正在尝试确定摄影疗法会导致免疫原性死亡，这可以使免疫系统启动（AIM 5）。在免疫能力的小鼠中确认了我们的发现后，我们将与免疫疗法（PD-1/PD-L1治疗 - AIM; AIM; 4）结合使用摄影疗法（单抗体 - 药物结合物和多种抗体 - 毒物结合物）。最后，我们希望与NCI或商业合作者合作，在I期临床试验中将这种治疗引入患者。

项目成果

Bilateral Ureteroenteric Strictures: A Case of the "Reverse 7".

双侧输尿管肠狭窄：“反向7”一例。

• DOI: 10.1016/j.urology.2018.04.022
• 发表时间: 2018
• 期刊: Urology
• 影响因子: 2.1
• 作者: Rayn,KareemN;Ritchie,Cayde;Folio,LesR;Stamatakis,Lambros;Verghese,MohanM;Agarwal,PiyushK
• 通讯作者: Agarwal,PiyushK

Clinical Trials Corner.

临床试验角。

• DOI: 10.3233/blc-189028
• 发表时间: 2018
• 期刊: Bladder cancer (Amsterdam, Netherlands)
• 作者: Agarwal,PiyushK;Sternberg,CoraN
• 通讯作者: Sternberg,CoraN