Localized immunotherapy using alum-binding therapeutics

项目摘要/摘要 旨在刺激协同免疫途径的联合治疗，使用细胞因子或 在临床前癌症模型中，免疫调节抗体通常比单一疗法更有效 免疫疗法。然而，当系统地给予这些联合治疗时，这些联合治疗遭受以下高毒性 靶上、肿瘤外刺激以及由于肿瘤穿透性差而在肿瘤部位的低局部浓度 以及很高的通过率。局部肿瘤内治疗是绕过某些挑战的可行方法。 与全身给药有关，但需要优化以促进治疗剂在 注射部位，并最大限度地减少泄漏进入循环。我们最近开发了一种方法来增强 通过改变免疫原与常用的氢氧化铝佐剂的结合来实现疫苗效力 (明矾)通过位点特异性的磷酸丝氨酸(PSer)多肽标签。PSer部分经历了配体交换 与明矾表面自由羟基的反应使蛋白质稳定地锚定在明矾上 粒子。我们建议在癌症的背景下应用这种明胶锚定平台来保持强大的免疫力 肿瘤部位内的激动剂，以最小的毒性促进强大的全身免疫反应。我们的 初步结果表明，这种简单的方法可以用来将刺激性细胞因子负载到明矾上，以 在肿瘤部位滞留长达一个月，刺激单针治疗产生强烈的抗肿瘤反应。 我们计划通过以下具体目标开发和优化这一翻译策略：(1)使用In-cell 磷酸化以产生磷酸丝氨酸标记的细胞因子和其他候选免疫激动剂以实现最佳 明胶结合，(2)确定这些肿瘤内明胶结合的治疗药物的最佳治疗方案 在多种肿瘤模型中的活体，(3)定义该疗法引起反应的作用机制， (4)评估全身免疫反应，并评估通过促进 将免疫刺激有效载荷转移到可移动的淋巴细胞，以运输到远端未经治疗的肿瘤。 这些研究将建立一个强大的技术平台，能够安全地提供目前看到的治疗方法 通过解决现有局部治疗策略中的关键限制，

Project Summary/Abstract Combination treatments aiming to stimulate synergistic immune pathways employing cytokines or immunomodulatory antibodies are generally more effective than monotherapies in preclinical models of cancer immunotherapy. However when given systemically, these combination treatments suffer from high toxicity from on-target off-tumor stimulation as well as low local concentrations at the tumor site due to poor tumor penetrance and high clearance rates. Local intratumoral therapy is a viable approach to bypass some of the challenges associated with systemic delivery, but requires optimization to promote retention of the therapeutic agent at the injection site and minimize leakage into the circulation. We have recently developed an approach to enhance vaccine efficacy by engineering the binding of immunogens to the commonly used adjuvant aluminum hydroxide (alum) via a site-specific phosphoserine (pSer) peptide tag. The pSer moieties undergo a ligand-exchange reaction with free hydroxyl groups on the surface of alum leading to stable anchoring of proteins on alum particles. We propose here to apply this alum-anchoring platform in the context of cancer to retain potent immune agonists within the tumor site, promoting a robust systemic immune response with minimal toxicity. Our preliminary results show that this simple approach can be used to load stimulatory cytokines onto alum for retention at the tumor site up to a month, stimulating a strong anti-tumor response from a single shot treatment. We plan to develop and optimize this translational strategy through the following specific aims: (1) use in-cell phosphorylation to produce phosphoserine-tagged cytokines and other candidate immune agonists for optimal alum binding, (2) determine optimal treatment regimens for these intratumoral alum-bound therapeutic agents in vivo in multiple tumor models, (3) define the mechanism of action through which this therapy elicits a response, (4) evaluate the systemic immune response and assess strategies to enhance abscopal effects by promoting the transfer of immunostimulatory payloads to motile lymphocytes for trafficking to distal untreated tumors. These studies will establish a robust technology platform capable of safely delivering treatments currently viewed as too toxic, by addressing key limitations in existing localized therapeutic strategies