Tackling the MARCKS-PIP3 Circuit to Attenuate Chronic Pulmonary Fibrosis

项目摘要 肺纤维化是正常肺损伤修复过程中的重要环节。然而，失控的伤害和修复， 而肺实质胶原过度沉积是慢性肺病的病理特征。 纤维化，如特发性肺纤维化(IPF)。这种疾病的中位生存期只有3到5年。 从确诊之日起数年。目前，除了两种药物外，还没有合适的药物用于治疗： FDA批准的九替丹尼和吡非尼酮。然而，不利和偏离目标的影响，以及未能 证明接受治疗的患者寿命延长表明迫切需要新的更好的治疗方法 代理人(S)治疗这种毁灭性的疾病。上皮间充质(EM)和成纤维细胞-肌成纤维细胞(FM) 过渡期与纤维化肺发病机制的启动和进展有关。新兴市场和 FM转变现象是与癌症恶性相关的重要致病事件，最初和 主要由受体介导的酪氨酸激酶(RTK)和PI3K-AKT信号通路介导。我们已经展示了 与EM转化相关的肺癌组织/细胞中磷酸化MARCKs的升高及其应用 多肽抑制剂MPS(Marcks PSD/ED序列)抑制EM转化和肺癌 通过处理与癌症发病机制相关的Marcks-PIP3通路的异常来治疗恶性肿瘤。这个 在组织切片和分离的成纤维细胞中也可以看到磷酸化Marcks升高的现象 IPF肺，但在任何正常的非纤维性肺中未见。我们最近的出版物显示了这种治疗方法 MPS多肽在抑制博莱霉素诱导的小鼠肺纤维化中的作用。在……里面 在体外，MPS处理异常的Marcks-PIP3回路以抑制Marcks的磷酸化，并选择性地 抑制EM/FM转化和肌成纤维细胞的纤维形成，以及M1/M2巨噬细胞的改变 极化。这种选择性只发生在IPF来源的成纤维细胞和活化的巨噬细胞上，而不是在 正常和灭活单核细胞。通过对多肽的优化，我们进一步开发出了稳定、更 生物安全性，以及一种新的MPS衍生肽MPS-6413DTM的高效性。初步研究已经证明了这种方法的有效性。 该多肽对博莱霉素诱导的小鼠肺纤维化病变的抑制和死亡作用，但对小鼠肺纤维化无明显影响 控制组的人。我们推测MPS-6413D是一种有效的抗肺纤维化药物，具有抑制纤维化的作用。 通过Marcks-PIP3通路处理慢性肺纤维化的进展。为了检验这一假设和 对于该多肽的治疗潜力，提出了两个目标。目的1是进一步确定治疗效果。 MPS-6413D多肽对32周龄(相当于42岁)博莱霉素性肺纤维化的治疗作用 老人)老鼠。其疗效将取决于对纤维原蛋白表达的抑制。 标记蛋白及其RNA，以及对总肺组织匀浆中Marcks及其磷酸化的抑制， 在这些年龄段的小鼠中，基质沉积、肺功能以及总体存活和幸福感 接触博莱霉素。目的2评价MPS-6413D多肽对肺间质纤维化的抑制作用 人源化小鼠模型中的肺成纤维细胞。为了更好地反映临床情况，IPF人肺成纤维细胞 (HLFS)将过继转移到C.B-17 SCID/BG小鼠体内，以测试MPS在体内的治疗潜力 肺纤维化。这些研究的成功将导致第二阶段SBIR动物研究的提交 自发性肺纤维化，药效学/药代动力学分析，FDA的一项基于IND的研究 未来临床药物开发以减轻慢性肺纤维化。

Project Summary Lung fibrosis is an important step of normal lung injury-repair process. However, uncontrolled injury and repair, and excessive deposition of collagen in the lung parenchyma is the pathological hallmark of chronic pulmonary fibrosis, such as idiopathic pulmonary fibrosis (IPF). The disease exhibits a median survival time of only 3 to 5 years from the time of diagnosis. Currently, there is no suitable drug for the treatment, except two drugs: Nintedanib and Pirfenidone, approved by FDA. However, adverse and off-target effects, and failure to demonstrate increased longevity in treated patients indicate the urgent need for new and better therapeutic agent(s) to treat this devastating disease. Epithelial-mesenchymal (EM) and fibroblast-myofibroblast (FM) transitions have been implicated in the initiation and the progression of fibrotic lung pathogenesis. The EM and FM transition phenomena, important pathogenic events associated with cancer malignancy, are primordially and mainly mediated by receptor-mediated tyrosine kinase (RTK) and PI3K-AKT signaling pathways. We have shown before the elevation of phospho-MARCKS in lung cancer tissues/cells associated with EM transition and the use of a peptide inhibitor, MPS (MARCKS PSD/ED Sequence), to suppress EM transition and lung cancer malignancy through tackling the aberrant MARCKS-PIP3 circuit associated with cancer pathogenesis. The elevated phospho-MARCKS phenomenon is also seen in tissue sections and isolated fibroblasts derived from IPF lungs, but not seen in any normal, non-fiberotic ones. Our recent publication had shown the therapeutic potential of MPS peptide in the suppression of the fibrotic lesions in bleomycin-induced fibrotic mouse lungs. In vitro, MPS tackles the aberrant MARCKS-PIP3 circuit to suppress MARCKS phosphorylation and also selectively inhibits the EM/FM transition and myofibroblast fibrogenesis, as well as the alteration of M1/M2 macrophage polarization. The selectivity occurs only on IPF-derived fibroblasts and activated macrophage, but not on the normal and inactivated monocytes. Through peptide optimization, we have developed further a stable, more biosafe, and high potency of a novel MPS-derived peptide, MPS-6413DTM. Initial studies have shown the efficacy of this peptide on the suppression of bleomycin-induced lung fibrotic lesions and deceased in mice, but not on the control ones. We hypothesize that MPS-6413D is a potent anti-fibrotic lung drug on the inhibition of fibrogenic progression of chronic lung fibrosis through tackling the MARCKS-PIP3 circuit. To test this hypothesis and the therapeutic potential of this peptide, two aims are proposed. Aim 1 is to determine further the therapeutic effects of MPS-6413D peptide on bleomycin induced lung fibrotic lesions in aged 32-week old (equivalent to 42 years old human) mice. The therapeutic potency will be determined on the inhibition of the expression of profibrogenic marker proteins and their RNA, and the inhibition of MARCKS and its phosphorylation in total lung homogenates, matrix deposition, pulmonary function, and also the overall survival and well-being in these age mice after bleomycin exposure. Aim 2 is to evaluate an inhibitory effect of MPS-6413D peptide on fibrogenic activity of IPF lung fibroblasts in humanized mouse model. To better reflect clinical scenarios, IPF human lung fibroblasts (HLFs) will be adoptively transferred to C.B-17 SCID/bg mice in order to test the therapeutic potential of MPS in lung fibrosis. Success of these studies will lead to the submission of the Phase II SBIR study on animals with spontaneous lung fibrosis, the pharmacodynamic/pharmacokinetic analysis, an IND-based study to FDA for future clinical drug development to attenuate chronic pulmonary fibrosis.