Localized immunotherapy using alum-binding therapeutics

项目概要/摘要 旨在利用细胞因子或细胞因子刺激协同免疫途径的联合治疗 在癌症的临床前模型中，免疫调节抗体通常比单一疗法更有效 免疫疗法然而，当全身给予时，这些组合治疗具有高毒性， 靶向肿瘤外刺激以及由于肿瘤转移率差而导致的肿瘤部位局部浓度低 和高清除率。局部瘤内治疗是绕过一些挑战的可行方法 与全身递送相关，但需要优化以促进治疗剂在全身递送中的保留。 注射部位，并尽量减少泄漏到循环。我们最近开发了一种方法， 通过工程化免疫原与常用佐剂氢氧化铝的结合来提高疫苗效力 （明矾）通过位点特异性磷酸丝氨酸（pSer）肽标签。pSer部分进行配体交换 与明矾表面上的游离羟基反应，导致蛋白质稳定锚定在明矾上 粒子我们在这里提出将这种明矾锚定平台应用于癌症的背景下，以保留有效的免疫原性。 在肿瘤部位内的激动剂，以最小的毒性促进稳健的全身免疫应答。我们 初步结果表明，这种简单的方法可用于将刺激性细胞因子加载到明矾上， 保留在肿瘤部位长达一个月，从单次注射治疗中刺激强烈的抗肿瘤反应。 我们计划通过以下具体目标来开发和优化这种翻译策略：（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，用于最佳免疫调节。 明矾结合，（2）确定这些肿瘤内明矾结合治疗剂的最佳治疗方案， 在多种肿瘤模型中的体内，（3）确定该疗法引起应答的作用机制， (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运动淋巴细胞，以运输到远端未治疗的肿瘤。 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所看到的治疗方法。 毒性太大，通过解决现有局部治疗策略的关键局限性，

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