Regeneration and remodeling of collecting lymphatic vessels post-injury

摘要 许多外科手术的侮辱--如淋巴清扫和淋巴活检--可能会对 淋巴管，特别是与放射治疗相结合时，会导致长期淋巴功能障碍 和淋巴水肿的形成。了解手术后淋巴再生的机制和 癌症治疗是预防淋巴水肿和开发有效的淋巴水肿治疗方法的关键。尽管 淋巴水肿的流行，尚不清楚收集血管是否在损伤后重建/再生 这种生长和重塑反应的机制尚不清楚。淋巴网络的破坏会导致 在剩余的网络中持续的液体负荷导致淋巴功能障碍。不适当的抵押和 收集因持续负荷而导致的血管重塑可导致不适应的重塑反应。在这里我瞄准了 发展对淋巴管重塑、修复和功能障碍的理解 使用复杂的淋巴成像、复杂的老鼠模型和计算建模的网络。这就做 研究淋巴流驱动淋巴重塑和再生的机制和模式。这就做 检测集合淋巴管在损伤后是否再生/重塑，并确定淋巴流动的影响 关于这一回应(目标1)。我还将测试淋巴流量改变和 利用遗传小鼠模型激活淋巴管生成通路。此外，我将确定 损伤部位的侧枝生长与小鼠的网络中断和淋巴流动改变有关 模型(目标2)。我还将使用3D培养系统来研究先前存在的附带增长的动态 收集容器在体外的流动和压力条件下。计算建模将补充目标2到 更好地了解结构重塑与淋巴流动动力学改变的关系。使用 在这个项目完成后，我将提供对预防策略和更好的治疗干预措施的见解 用于淋巴浮肿治疗。 我在淋巴生物工程，淋巴成像，图像处理，分离淋巴方面有广泛的培训 从我的博士训练中学到的容器实验和计算模型。该项目将利用此培训 在我的博士后导师-帕德拉博士和穆恩博士-的实验室里，他们是淋巴学领域的领军人物 研究和生物学。帕德拉博士的实验室已经开发出最先进的淋巴成像工具，可以精确地 测量淋巴流率、淋巴结动态活体显微镜和淋巴动物模型 疾病。穆恩博士的实验室开发了血管生成的体外模型，即血管生成的计算模型 以及淋巴运输、生物工程细胞培养系统和跟踪细胞的图像处理工具。这 独特的培训环境将使我能够成功地完成这项建议的目标。

SUMMARY Many surgical insults—such as lymph node dissection and lymph node biopsy —can cause damage to the lymphatic vessels, particularly when combined with radiotherapy, resulting in long-term lymphatic dysfunction and the formation of lymphedema. Understanding the mechanisms of lymphatic regeneration after surgery and cancer treatment is crucial to prevent lymphedema and develop efficient lymphedema therapy. Despite the prevalence of lymphedema, it is not clear whether collecting vessels remodel/regenerate post-injury and mechanisms of such growth and remodeling response are unknown. A disrupted lymphatic network induces sustained fluid loads in the remaining network that drives lymphatic dysfunction. Improper collateralization and collecting vessel remodeling due to sustained loads can cause a maladaptive remodeling response. Here I aim to develop an understanding of lymphatic vessel remodeling, repair and dysfunction in the context of a disrupted network using sophisticated lymphatic imaging, complex mouse models, and computational modeling. I will investigate the mechanisms and modes by which lymph flow drives lymphatic remodeling and regrowth. I will test whether collecting lymphatic vessels regrow/remodel following injury and determine the effect of lymph flow on this response (Aim 1). I will also test whether there is a causal relationship between altered lymph flow and activation of lymphangiogenic pathways using genetic mouse models. Further, I will determine the dynamics of collateral growth at the injury site in the context of a disrupted network and altered lymph flow in our mouse model (Aim 2). I will also use a 3D culture system to study the dynamics of collateral growth from pre-existing collecting vessels under flow and pressure conditions in vitro. Computational modeling will complement Aim 2 to better understand the relationship between structural remodeling and alteration of lymph flow dynamics. With the completion of this project, I will provide insight into preventive strategies and better therapeutic interventions for lymphedema therapy. I have extensive training in lymphatic bioengineering, lymphatic imaging, image processing, isolated lymphatic vessel experiments and computational modeling from my doctoral training. This project will leverage this training in the labs of my postdoctoral advisors—Dr. Padera and Dr. Munn—who are leaders in the field of lymphatic research and biology. Dr Padera’s lab has developed state-of-the-art lymphatic imaging tools to precisely measure lymph flow rate, dynamic intravital microscopy of lymph nodes and animal models of lymphatic diseases. Dr Munn’s lab has developed in vitro models of angiogenesis, computational models of angiogenesis and lymphatic transport, bioengineered cell-culture systems and image processing tools to track cells. This unique training environment will allow me to successfully complete the aims of this proposal.