Administration of intratumoral immunocytokine to activate immune rejection of spontaneous canine melanoma

恶性黑色素瘤是一个严重的健康问题，退伍军人，转移性黑色素瘤通常是无法治愈的，如果它是 转移到远处。免疫检查点阻断的令人兴奋和令人印象深刻的数据表明， 免疫系统在一些转移性黑色素瘤患者中产生持久应答的能力， 改变了护理标准1。免疫检查点阻断的有效治疗似乎需要 激活对多种黑素瘤抗原特异的抗黑素瘤T细胞，包括患者- 独特的新抗原犬恶性黑色素瘤为研究黑色素瘤提供了极好的临床前模型 免疫治疗，因为它类似于在完整免疫环境中自发发生的人黑色素瘤。 系统和通过淋巴管或血管转移到区域淋巴结，肺，肝， 大脑和肾脏根据本申请，GD 2二唾液酸神经节苷脂（GD 2）在两种人类中表达， 和犬黑色素瘤2 -4因此，我们建议瘤内（IT）注射GD 2反应性hu14.18-IL 2 免疫细胞因子（IC）（IT-IC）单独和与其他疗法联合在临床前小鼠中的协同作用 模型，在患有黑素瘤的伴侣犬中将注射的肿瘤转化为有效的原位肿瘤 疫苗5.我们的中心假设是IT-IC结合局部放射治疗（RT）在犬中 黑色素瘤可以诱导针对黑色素瘤的T细胞应答。此外，我们假设这种反应可以是 我们认为，该方案是安全的，并且耐受性良好。这些 将通过实现以下目标来检验假设：目的1）确定毒性、免疫原性，并评估 在患有局部晚期或晚期乳腺癌的狗中，单独和与局部RT一起IT递送hu14.18-IL 2的抗肿瘤活性 转移性黑色素瘤;目的2）评估IT递送hu14.18后的局部和全身抗肿瘤活性。 IL 2联合RT和抗PD 1免疫检查点阻断治疗局部晚期或 转移性黑色素瘤;和目的3）鉴定生物标志物以告知与IT的组合免疫治疗策略 在患有局部晚期或转移性黑素瘤的狗中递送hu14.18-IL 2。标准临床评估 毒性和耐受性的评估以及临床明显疾病的测量和成像是目标1的一部分 和2.系列血液样本和肿瘤活检将允许详细的组织学和免疫学评估 在目标3中，确定抗肿瘤活性的机制并确定组织学发现是否 在我们的鼠模型中观察到的伴随免疫耐受也存在于狗中。探索性实验室研究 将：i）评估免疫治疗前后外周和肿瘤细胞环境的变化，ii） 利用新的免疫监测来鉴定患有黑素瘤的狗的应答的候选生物标志物 接受IT-IC，和iii）评估肿瘤内和外周血T细胞受体的多样性和克隆性 保留曲目。这项研究使用自发性犬黑色素瘤来测试一种新的RT + IT-IC +免疫检查点 阻断方案已被证明可诱导持久、强效和有效消除大的局部 和远处建立的荷瘤小鼠黑色素瘤病变5。这项犬类试验的结果 使这些概念能够在人类患者（包括退伍军人）中快速转化为临床测试， 晚期黑素瘤这种方法在临床上是有利的，因为它涉及的试剂/治疗是 目前可用的或在临床试验中，并且它可以容易地作为“现成”疗法提供， 美国各地的退伍军人医院。此外，它通过针对有效和患者的个性化治疗- 特异性新抗原。最后，RT + IT-IC +免疫检查点阻断可纳入治疗 几乎任何其他恶性肿瘤，包括退伍军人中的其他常见恶性肿瘤。唯一的要求 对于这种策略，具有针对所靶向的恶性肿瘤的肿瘤反应性单克隆抗体（mAb）， 类似于hu14.18，可以用公式表示为IC或IC存在。

Malignant melanoma is a serious health issue for Veterans, and metastatic melanoma is usually incurable if it metastasizes to distant sites. Exciting and impressive data with immune checkpoint blockade demonstrate the ability of the immune system to produce durable responses in some metastatic melanoma patients and have changed the standard of care1. Effective treatment with immune checkpoint blockade seems to require activation of anti-melanoma T cells that are specific for a wide variety of melanoma antigens including patient- unique neoantigens. Canine malignant melanoma provides an excellent preclinical model to study melanoma immunotherapy as it is similar to human melanoma occurring spontaneously in the setting of an intact immune system and with metastasis occurring via lymphatics or blood vessels to regional lymph nodes, lungs, liver, brain, and kidney. Germane to this application, the GD2 disialoganglioside (GD2) is expressed in both human and canine melanoma2-4. We therefore propose intratumoral (IT) injection of the GD2-reactive hu14.18-IL2 immunocytokine (IC) (IT-IC) alone, and in combination with other therapies synergistic in preclinical murine models, in companion dogs with melanoma to convert the injected tumor into an effective in situ tumor vaccine5. Our central hypothesis is that IT-IC in combination with local radiation therapy (RT) in canine melanoma can induce a T cell response to melanoma. Further, we hypothesize that this response can be amplified with immune checkpoint blockade, and that this regimen is safe and well tolerated. These hypotheses will be tested by achieving the following: Aim 1) Determine toxicity, immunogenicity, and assess antitumor activity of IT delivery of hu14.18-IL2 alone and with local RT in dogs with locally advanced or metastatic melanoma; Aim 2) Evaluate local and systemic antitumor activity following IT delivery of hu14.18- IL2 combined with RT and immune checkpoint blockade with anti-PD1 in dogs with locally advanced or metastatic melanoma; and Aim 3) Identify biomarkers to inform combination immunotherapy strategies with IT delivery of hu14.18-IL2 in dogs with locally advanced or metastatic melanoma. Standard clinical assessments of toxicity and tolerance, as well as measurement and imaging of clinically evident disease, are part of Aims 1 and 2. Serial blood samples and tumor biopsies will allow for detailed histologic and immunologic assessments in Aim 3 to determine mechanisms of antitumor activity and to determine whether histologic findings of concomitant immune tolerance seen in our murine model are also present in the dog. Exploratory lab studies will: i) evaluate alterations in the cellular milieu in the periphery and tumor before and after immunotherapy, ii) utilize novel immune monitoring to identify a candidate biomarker of response for dogs with melanoma receiving IT-IC, and iii) assess the diversity and clonality of intratumoral and peripheral blood T cell receptor repertoire. This study uses spontaneous canine melanoma to test a novel RT + IT-IC + immune checkpoint blockade regimen already proven to induce long lasting, potent and effective elimination of large local and distant established melanoma lesions in tumor-bearing mice5. The results from this canine trial could enable rapid translation into clinical testing of these concepts in human patients, including Veterans, with advanced melanoma. This approach is clinically advantageous as it involves agents/treatments that are currently available or in clinical testing, and it could be readily made available as an “off the shelf” therapy in Veterans Hospitals throughout the USA. Moreover, it personalizes treatment by targeting potent and patient- specific neoantigens. Finally, RT + IT-IC + immune checkpoint blockade could be incorporated into treatment of virtually any other malignancy, including other common malignancies in Veterans. The only requirement for this strategy is having a tumor-reactive monoclonal antibody (mAb) for the malignancy being targeted, analogous to hu14.18, that could either be formulated as an IC or for which an IC exists.