Safety Assessment of Laser Activated NanoTherapy (LANT) as a Potential Treatment for Veterans with Cutaneous Squamous Cell Carcinomas

皮肤鳞状细胞癌(CSCC)是皮肤癌的两倍多。 死亡人数超过黑色素瘤，占所有皮肤癌的50%。大约有70万个病例是 在美国每年确诊一次，但退伍军人患CSCC的风险更高，因为年龄和 在执勤期间大量接触紫外线辐射和含砷的化学战剂。多数 CSCC可以通过手术切除或Mohs显微外科手术(Mohs)成功地治疗；然而， 部分CSCC病例可导致局部复发、转移和死亡。对许多人来说，糟糕的前景预示着 患有CSCC的退伍军人对更好的治疗选择的需求大量未得到满足，特别是对于那些 要求非手术治疗方案或不适合手术的患者，因为他们 免疫抑制或有其他合并症。该提案中的新技术--激光激活 当手术不是一种选择时，纳米疗法(LANT)可以满足这一尚未满足的替代放射治疗的需求。 LANT是一种使用近红外激光激发金纳米棒(AuNRs)的热消融平台疗法 这会产生足够的热量来诱导特定部位的细胞死亡。我们的技术创新是 纳米颗粒设计和由此产生的治疗结果。在我们的初步研究中，兰特证明了 (A)在我们的体外剂量-升级曲线上，最高浓度的SCC细胞100%死亡，以及(B)完全死亡 皮下移植肿瘤边缘清晰的一次治疗后肿瘤消退 小鼠鳞状细胞癌，与三个对照组相比，未治疗组、仅激光组和AuNRs组- 只有这样。基于这些发现，我们提出了一项旨在证明LANT是安全的和可翻译的研究 兰特从长凳到床边，有三个目标。对于目标1，我们将演示生物分布、清除、 AuNRs在小鼠肿瘤模型和家猪模型中的安全性 获取主键/PD数据 使用电感耦合等离子体- MS、血液学/血清学和组织学。我们将验证AuNRs、发热量和LANT的分布- 使用K型三维超声成像对肿瘤、肿瘤边缘和周围组织造成的损伤 热电偶和红外热像仪。我们还将在不同的激光输出下验证LANT的安全性 通过摄影成像捕捉大体外观变化并分析形态的能力 随着肿瘤、肿瘤边缘和周围组织的组织学评估而变化。我们将与我们的 FDA顾问和七(7)个临床合作伙伴在开始试验之前获得FDA和IRB的批准 学习。对于目标2，我们将招募有活检证实的局部低风险CSCC的退伍军人(n=10)。退伍军人 将在LANT期间接受三维超声引导下的AuNRs注射。退伍军人也将获得这一标准 术后21天行Mohs显微外科手术以确认肿瘤边缘清楚。我们将聘请一名 自适应贝叶斯连续再评估方法(CRM)用于确定和调整LANT剂量， 用三维超声成像和数字化技术测量局部皮肤毒性和肿瘤消退 卡尺。对于目标3，我们将评估肿瘤的形态和组织病理学变化，肿瘤 LANT前后的切缘及周围组织。局灶性卵巢癌的组织病理学评价 活检和治疗的肿瘤将被用来表征黑素细胞的外观，表皮， 肿瘤的真皮、皮下组织、皮下组织、肿瘤边缘和周围组织在LANT手术前后。 我们将从生物学、形态学和组织病理学的角度深入了解LANT对细胞和肉眼的影响， 证明LANT是一种安全有效的治疗CSCC的方法。

Cutaneous squamous cell carcinomas (cSCC) are responsible for more than twice the number of skin cancer deaths than melanoma, representing up to 50% of all skin cancers. Approximately 700,000 cases are diagnosed annually in the United States, but Veterans have a higher risk for cSCC because of age and extensive exposures during the line of duty to UV radiation and arsenic-based chemical warfare agents. Most cSCC can be successfully treated by surgical excision or Mohs Micrographic Surgery (Mohs); however, a subset of cSCC cases result in locoregional recurrence, metastases, and death. The poor prognoses for many Veterans with cSCC result in a substantial unmet need for better treatment options, especially for those who request a non-surgical treatment option or who are not a candidate for surgery because they are immunosuppressed or have other comorbidities. The novel technology in this proposal, Laser-Activated NanoTherapy (LANT), can meet this unmet need for an alternative to radiation when surgery is not an option. LANT is a thermal ablation platform therapy using near-infrared laser excitation of gold nanorods (AuNRs) that generates enough heat to induce site-specific cellular death. The innovation of our technology is the nanoparticle design and the resulting therapeutic outcomes. In our preliminary studies, LANT demonstrated (a) 100% SCC cell death at the highest concentration on our dose-escalation curve in vitro, and (b) complete tumor regression after one treatment iteration with clear tumor margins in subcutaneously-xenografted squamous cell carcinoma in mice, compared to three control groups, no-treatment, laser-only, and AuNRs- only. Based on these findings, we propose a study designed to demonstrate that LANT is safe and translate LANT from bench to bedside with three aims. For Aim 1, we will demonstrate the biodistribution, clearance, and safety of AuNRs in a murine tumor model and a domestic swine model to acquire PK/PD data using ICP- MS, hematology/serology, and histology. We will verify the distribution of AuNRs, heat generation, and LANT- induced injury to the tumor, tumor margins, and surrounding tissues using 3D ultrasound imaging, K-type thermocouples, and IR thermographic cameras. We will also validate LANT safety at different laser output powers by capturing gross appearance changes with photographic imaging and analyzing morphological changes with histological evaluation of tumors, tumor margins, and surrounding tissues. We will work with our FDA consultant and seven (7) clinical partners to obtain FDA and IRB approvals before beginning the pilot study. For Aim 2, we will recruit Veterans (n = 10) who have biopsy-confirmed local, low-risk cSCC. Veterans will receive 3-D ultrasound-guided injections of AuNRs during LANT. Veterans will also receive the standard of care Mohs Micrographic Surgery to confirm clear tumor margins 21 days after LANT. We will employ an adaptive Bayesian Continual Reassessment Method (CRM) to determine and adjust the LANT dose, measured by local skin toxicities and tumor regression measured by 3-D ultrasound imaging and digital calipers. For Aim 3, we will evaluate the morphological and histopathological changes of the tumor, tumor margin, and surrounding tissue before and after LANT. Histopathological evaluation of the locoregional biopsies and treated tumors will be used to characterize the appearance of melanocytes, the epidermis, dermis, hypodermis, and subcutis of the tumor, tumor margin, and surrounding tissue before and after LANT. We will gain biological, morphological, and histopathological insight into LANT's cellular and gross impact, demonstrating that LANT is a safe and effective treatment option for cSCC.