Targeted Anti-IL-1β Platelet Mimetics for Cardiac Detoxification and Repair

项目摘要 目前，对心脏的缺血性损伤不能通过传统的医疗护理来修复，因此仅 存在姑息治疗。干细胞移植是治疗心脏病的一个有前途的策略 再生，但目前的疗法受到再生细胞和再生细胞之间相互作用不足的限制。 受伤的组织在上一个资助期，我们开发了靶向纳米颗粒（即双特异性抗体- 偶联剂）以将循环干细胞重定向至梗塞心脏用于治疗性再生。尽管 这样的初步成功，我们意识到我们的系统有一些问题：（P1）我们不能完全回答干细胞 （“种子”）用于心脏修复。损伤后的心脏微环境（“土壤”）需要准备好， 最大结果;（P2）抗体靶向非常特异，但完全依赖于抗原， 心脏损伤生物标志物仅在损伤后的短时间内表达。当前更新 该提案建立在以前的研究基础上，但在技术和 概念上。为了解决P1，我们认为其中一种抗体需要是治疗性的，以对抗P1。 心脏过度炎症。为了解决P2，我们寻找与P2具有广谱亲和力的药物。 心脏损伤为此，我们开发了抗IL-1血小板模拟物（IL-1-PM）。IL 1-PM的作用方式 如下：血小板囊泡作为我们系统的载体，它们具有天然的能力， 损伤（在血小板膜上的结合基序上应答）;抗IL-1抗体目前处于3期 临床试验，并已证明有能力中和炎症和促进心脏修复;血小板 囊泡可以进一步装载干细胞衍生的生长因子以帮助修复过程。目标1：制造 IL-1-PM，并表征其理化和生物学性质。我们将通过以下方式生成IL 1-PM代理 将抗IL-1抗体缀合到血小板膜纳米囊泡上;结合/接合能力，毒性， 将在培养的细胞和健康动物中检查IL 1-PM的药代动力学。目标2：确定 间充质干细胞（MSC）分泌组负载IL 1-PM在小鼠模型中的治疗潜力 心肌梗死心肌缺血再灌注可诱发心肌梗死。之后，MSC-IL 1-PM，沿着与各种 对照剂将通过静脉内递送。将确定治疗安全性和疗效。此外，本发明还提供了一种方法， 将探讨这种治疗的基本机制。目标3：将研究结果转化为临床- 相关的大型心肌梗死动物模型。将通过球囊闭塞在猪中诱导MI procedure.将评价MSC-IL 1-PM治疗的安全性和有效性。我们的治疗系统 联合干细胞治疗（组分1）和抗IL-1治疗（组分2），这两种治疗都已被 在心脏修复的临床试验中经过严格测试和验证。此外，治疗剂将以可接受的方式递送。 依赖于血小板结合基序的损伤发现能力的靶向方式（组分3）。所有3个组件 得到了我们小组强有力的初步数据的支持。

PROJECT SUMMARY Currently, ischemic damage to the heart cannot be repaired by conventional medical care therefore only palliative treatments exist. Stem cell transplantation is a promising strategy for therapeutic cardiac regeneration, but current therapies are limited by insufficient interaction between the regenerative cells and the injured tissue. In the last grant period, we have developed targeted nanoparticles (namely bispecific antibody- conjugated agents) to redirect circulating stem cells to the infarcted heart for therapeutic regeneration. Despite such initial success, we realize our system has some problems: (P1) We cannot fully reply on the stem cells (“seeds”) for cardiac repair. The post-injury heart microenvironment (“soil”) needs to be primed for the maximum outcome; (P2) Antibody targeting is quite specific but is fully dependent on the antigen, which are cardiac injury biomarkers that only expresses in a short period of time after injury. The current renewal proposal builds on the previous study, but represents a significant advancement, both technically and conceptually. To address P1, we reason one of the antibodies needs to be therapeutic, to combat the excessive inflammation in the heart. To address P2, we seek for agents that have broad spectrum affinity with cardiac injury. To those ends, we developed anti-IL-1 platelet mimetic (IL1-PM). The mode of action for IL1-PM is as follows: platelet vesicles serve as the carrier of our system and they have innate ability to find cardiac injury (replying on the binding motifs on platelet membranes); anti-IL-1 antibodies are currently in Phase 3 clinical trials and have demonstrated ability to neutralize inflammation and promote cardiac repair; platelet vesicles can be further loaded with stem cell-derived growth factors to aid the repair process. AIM 1: Fabricate IL1-PM and characterize its physicochemical and biological properties. We will generate IL1-PM agents by conjugating anti IL-1 antibodies onto platelet membrane nanovesicles; binding/engaging ability, toxicity, pharmacokinetics of IL1-PM will be examined in cultured cells and in healthy animals. AIM 2: Determine the therapeutic potential of mesenchymal stem cell (MSC) secretome-loaded IL1-PM in a mouse model of myocardial infarction. MI will be induced by ischemia-reperfusion. After that, MSC-IL1-PM, along with various control agents, will be delivered intravenously. Therapeutic safety and efficacy will be determined. In addition, the underlying mechanisms of such treatment will be explored. AIM 3: Translate the findings into a clinically- relevant large animal model of myocardial infarction. MI will be induced in swine via a balloon-occlusion procedure. The safety and efficacy of MSC-IL1-PM treatment will be evaluated. Our therapeutic system combines stem cell therapy (component 1) and anti-IL1 therapy (component 2), both of which have been rigorously tested and verified in clinical trials for cardiac repair. Moreover, the therapeutics will be delivered in a targetable fashion relying on the injury-finding ability of platelet binding motifs (component 3). All 3 components have been supported by strong preliminary data from our group.