Investigating the impact of tumor heterogeneity on sustained low dosage immunotherapy via a peritumoral immunotheranostic hydrogel

癌症治疗最具挑战性的方面之一是每个患者疾病的独特性，这可能导致有效诊断和治疗的严重延误。这个问题对于免疫疗法来说尤为重要，免疫疗法是一种使用治疗方法（药物）来修改或增强患者自身免疫系统靶向和杀死癌细胞的能力的疗法。目前的免疫治疗方法使用一系列高剂量的输注，例如将治疗药物注射到血液循环中，这可能严重破坏免疫系统的正常功能，导致严重且往往不可预测的副作用。对于黑色素瘤（皮肤癌）和其他实体瘤患者，对治疗的反应取决于肿瘤内细胞的独特种类和治疗药物到达这些细胞和组织的能力。因此，该项目的研究目标是设计一种可定制的递送系统，用于研究低剂量治疗策略，并评估不同治疗方法到达肿瘤内关键免疫细胞和癌细胞的能力。研究人员将通过测量两种黑色素瘤小鼠模型的肿瘤和细胞内治疗药物的积累来优化和验证该系统，这两种模型代表了由于不同肿瘤结构导致的两种患者特异性疾病的差异。与一系列高剂量输注不同，这些疗法将以可控的低剂量连续方式施用，以研究持续和局部的免疫治疗。这项工作可能会导致一种新的免疫疗法治疗选择，其副作用较少。该项目的教育目的是让本科生和公众了解生物医学工程在癌症研究领域的贡献。为本科生开设一门癌症工程课程，讨论基于基础工程的癌症治疗策略。课堂上的学生将维护一个相关的网站，重点教育公众工程学概念如何增强癌症治疗策略。本项目的目标是设计和应用一个新的系统来研究肿瘤异质性在持续低剂量免疫调节过程中对肿瘤微环境病理生理的影响。该项目的第一个目标是定制丝束（FM）水凝胶，用于持续的肿瘤周围免疫治疗（联合免疫治疗和诊断）胶束（MCs）和生物制剂的递送。1)通过DyLight 650和Gd（钆）标记的fm水凝胶，可通过磁共振成像（MRI）实时诊断评估肿瘤边缘异质性；2)持续低剂量递送免疫刺激剂（IMQ（咪喹莫特））以诱导髓源性抑制细胞（MDSCs）的促炎刺激和肿瘤驻留树突状细胞（dc）的细胞因子IL-12的局部释放；3)通过将IL-12加载到共价保留在fp水凝胶中的聚合体（PS）中直接持续低剂量递送IL-12。在体外研究中，光氧化将用于触发柱体到球体，即fm到mc，转变和释放PS有效载荷。研究旨在验证假设，即IMQ和Gd将从fm水凝胶转移到释放的胶束中，并在柱向球的转变过程中保持活性。该项目的第二个目标是研究免疫治疗性MCs和IL-12在体内局部和持续递送对临床相关异质性实体瘤微环境的影响。由于两种黑色素瘤小鼠模型（BRAF(V600E)/PTEN和B16F10）在血管发育和微环境中存在免疫抑制细胞方面存在显着差异，因此在策略上选择了它们。将绘制和比较淋巴引流的差异，以深入了解肿瘤边缘的异质性如何影响肿瘤内治疗的靶向性，并研究纳米材料与免疫细胞之间的相互作用，这对癌症免疫治疗至关重要。研究旨在验证两个假设：1)模型之间肿瘤异质性的差异将影响淋巴引流和胶束进入肿瘤微环境；2)与高剂量间歇注射相比，通过实体瘤的引流淋巴持续低剂量递送免疫调节因子将更有效地抑制肿瘤诱导的免疫抑制。除了有可能产生治疗和监测肿瘤内部变化的新方法外，该平台还可用于研究持续治疗给药方案，评估患者特异性血管和淋巴进入肿瘤的途径以进行个性化癌症治疗，以及研究特异性免疫调节因子对实体肿瘤进展和生长的影响。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

One of the most challenging aspects of cancer therapy is the uniqueness of each patient's disease, which can result in significant delays in effective diagnosis and treatment. This issue has become particularly critical for immunotherapy, which is therapy that uses therapeutics (drugs) to modify or enhance a patient's own immune system's ability to target and kill cancerous cells. Current immunotherapy methods use a series of high dose infusions, e.g., injections, of therapeutics into the blood circulation, which can severely disrupt the normal function of the immune system, resulting in harsh and often unpredictable side effects. For patients with melanoma (skin cancer) and other solid tumors, responses to treatment depend on both the unique variety of cells within the tumors and the therapeutics' ability to reach these cells and tissues. Thus, the research objective of this project is to engineer a customizable delivery system for investigating lower dose treatment strategies and for assessing the ability of different therapeutics to reach critical immune and cancer cells within tumors. The investigators will optimize and validate this system by measuring the accumulation of therapeutics within tumors and cells of two mouse models of melanoma, which are representative of two patient-specific disease differences due to different tumor structures. As opposed to a series of high dose infusions, these therapeutics will be administered in a controlled continuous fashion at low doses to investigate sustained and localized immunotherapy. This work may lead to a new immunotherapy treatment option with a lower number of unwanted side effects. The educational objective of this project is to expose undergraduate students and the public to the contributions of Biomedical Engineering in the field of cancer research. A Cancer Engineering course for undergraduates will be implemented to discuss fundamental engineering-based strategies to treat cancer. An accompanying website will be maintained by the students in the class that focuses on educating the public on how engineering concepts enhance strategies for cancer therapy.The goal of this project is to design and apply a novel system to investigate the impact of tumor heterogeneity on the pathophysiology of the tumor microenvironment during sustained low dosage immunomodulation. The project's first objective is to tailor filomicelle (FM)-hydrogels for the sustained peritumoral delivery of immunotheranostic (combined immunotherapy and diagnostics) micelles (MCs) and biologics. FM-hydrogels will be customized to function as a tool for 1) real time diagnostic assessment of tumor margin heterogeneity via magnetic resonance imaging ( MRI), enabled by a DyLight 650 and Gd (gadolinium) tagged FM-hydrogels; 2) the sustained low dosage delivery of an immunostimulant (IMQ (imiquimod)) to induce pro-inflammatory stimulation of myeloid-derived suppressor cells (MDSCs) and localized release of cytokine IL-12 from tumor resident dendritic cells (DCs) and 3) the sustained low dosage delivery of IL-12 directly by loading the IL-12 within polymersomes (PS ) that will be covalently retained within the FM-hydrogel. During in vitro studies, photo-oxidation will be used to trigger the cylinder-to-sphere, i.e., FM-to-MC, transition and release of PS payloads on demand. Studies were designed to test the hypothesis that both IMQ and Gd will transfer from the FM-hydrogels to released micelles during the cylinder-to-sphere transition and remain active. The project's second objective is to investigate the influence of in vivo localized and sustained delivery of immunotheranostic MCs and IL-12 to the microenvironment of clinically relevant heterogeneous solid tumors. Two melanoma mouse models (BRAF(V600E)/PTEN and B16F10) were strategically selected due to their significant differences in vascular development and presence of immunosuppressive cells within their microenvironments. The differences in lymphatic drainage will be mapped and compared to provide insight into how heterogeneity at the tumor margin can impact intratumoral targeting of therapeutics and the interactions between nanomaterials and immune cells critical to cancer immunotherapy will be examined. Studies were designed to test two hypotheses: 1) that differences in tumor heterogeneity between the models will impact lymphatic drainage and access of micelles to the tumor microenvironment and 2) that sustained low dosage delivery of immunomodulatory factors through the draining lymphatics of solid tumors will more efficiently inhibit tumor-induced immune suppression compared to high dosage intermittent injections. In addition to the potential to generate new approaches to treat and monitor changes within tumors, the platform developed may be useful for investigating sustained therapeutic dosing regimens, for assessing patient-specific vascular and lymphatic access to tumors for personalized cancer therapy and for investigating the impact of specific immunomodulatory factors on the progression and growth of solid tumors.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.