Lymphatic Regeneration by Direct Cellular Reprogramming

通过直接细胞重编程实现淋巴再生

基本信息

批准号：
10744935
负责人：
Sang-Ho Lee
金额：
$ 52.09万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10744935
关键词：
Address Adenovirus Vector Animal Model Biological Biological Response Modifier Therapy Body Fluids Bypass Cardiovascular Diseases Cardiovascular system Cell Lineage Cell Reprogramming Cell Therapy Cells Characteristics Clinical Data Dependence Development Disease Embryo Endothelial Cells Exhibits Fibroblasts Fluid Balance Generations Genes Genomics Goals Growth Histologic Human Imaging technology Immunologic Surveillance Impairment Incidence Insertion Mutation Lentivirus Vector Lymphatic Lymphatic Endothelial Cells Lymphatic function Lymphedema Magnetic Resonance Imaging Measurement Methods Molecular Morbidity - disease rate Mus Natural regeneration Near-infrared optical imaging Obstruction Palliative Surgery Play Regulation Research Residual state Role Somatic Cell Therapeutic Therapeutic Effect Tissues Transplantation Tumorigenicity Undifferentiated Variant Viral adult stem cell cell type clinical application clinical translation cost delivery vehicle experimental study gene therapy human embryonic stem cell human pluripotent stem cell in vivo induced pluripotent stem cell innovative technologies insight lymphatic development lymphatic vessel mortality neovascularization new therapeutic target novel novel strategies novel therapeutic intervention novel therapeutics overexpression paracrine programs regenerative therapy side effect stem cells success transcription factor vector wound healing

项目摘要

Project Summary (Abstract) Lymphatic vessels play an important role in tissue fluid homeostasis and immune surveillance. Thus, impaired function of lymphatic vessels due to abnormal vessel development or damaged lymphatic vessels causes obstruction of draining body fluid and leads to the development of lymphedema. Despite a continuous increase in the incidence of lymphedema, therapeutic options are limited to conservative treatment or palliative surgery. Over the past decades, new biological treatments such as gene or cell therapy have emerged for treating various cardiovascular diseases. However, the effects of adult stem cells turned out to be minimal, and embryonic or induced pluripotent stem cell (ESC/iPSC)-derived cells are costly to produce and maintain and may cause side effects. To avoid these problems, a new approach, called direct reprogramming or direct conversion has been developed, in which somatic cells are converted into other lineage cells by overexpression of lineage- or cell-type specific transcription factors (TFs). This approach allows simpler and safer target cell generation and has the potential for more convenient clinical translation. With this direct reprogramming approach, we have successfully generated reprogrammed endothelial cells (rECs) via ETV2, and more recently, reprogrammed lymphatic ECs (rLECs). The overall goal of this project is to develop a direct reprogramming approach for treating lymphedema. The direct EC reprogramming approach for therapy has two options: cell therapy or direct in vivo reprogramming. In this proposal, we will follow both options. For clinical application, both require a safer delivery vector to minimize the possibility of genomic integration. Thus, we developed an adenoviral-ETV2 (Ad-ETV2) vector. As a first approach, we will develop a cell-based therapy using rLECs. As a second approach, we will develop a direct in vivo reprogramming approach without using cells. In the latter, Ad-ETV2 will be injected into animal models to see whether host cells can be reprogrammed into LECs and form new lymphatic vessels. To date, none of these two approaches have been attempted for treating lymphedema. More specifically, in Aim 1, we will generate early and late rLECs from human fibroblasts by changing various culture conditions. In Aim 2, we will determine the therapeutic effects of rLECs on experimental lymphedema and the mechanisms underlying these therapeutic effects. In Aim 3, we will explore whether ETV2 is able to directly reprogram fibroblasts into functional LECs, induce lymphatic vessel formation, and ameliorate lymphedema. We anticipate that the results of the proposed experiments will yield new insight into the role of novel cell and gene therapy for treating lymphedema.

项目概要（摘要）淋巴管在组织液稳态和免疫监视中发挥重要作用。因此，由于血管发育异常或淋巴管受损而导致淋巴管功能受损导致体液排出受阻并导致淋巴水肿的发生。尽管连续淋巴水肿发生率增加，治疗选择仅限于保守治疗或姑息治疗外科手术。在过去的几十年里，新的生物疗法如基因或细胞疗法已经出现治疗各种心血管疾病。然而，成体干细胞的作用被证明是微乎其微的，并且胚胎或诱导多能干细胞 (ESC/iPSC) 衍生细胞的生产和维护成本高昂，可能会引起副作用。为了避免这些问题，一种新方法被称为直接重编程或直接已经开发出转化，其中体细胞通过过度表达转化为其他谱系细胞谱系或细胞类型特异性转录因子 (TF)。这种方法允许更简单、更安全的靶细胞生成并具有更方便的临床转化的潜力。通过这种直接重新编程方法，我们已经通过 ETV2 成功生成了重编程内皮细胞 (rEC)，最近，重新编程的淋巴 EC（rLEC）。该项目的总体目标是开发一种治疗淋巴水肿的直接重编程方法。直接内皮细胞重编程治疗方法有两种选择：细胞疗法或直接体内重编程。在本提案中，我们将遵循这两种选择。对于临床应用，两者都需要更安全的递送载体最大限度地减少基因组整合的可能性。因此，我们开发了腺病毒-ETV2 (Ad-ETV2) 载体。作为第一种方法是，我们将开发一种使用 rLEC 的细胞疗法。作为第二种方法，我们将开发一个不使用细胞的直接体内重编程方法。在后者中，Ad-ETV2将被注射到动物体内建立模型来观察宿主细胞是否可以被重编程为 LEC 并形成新的淋巴管。迄今为止，这两种方法均未尝试用于治疗淋巴水肿。更具体地说，在目标 1 中，我们通过改变各种培养条件，将从人类成纤维细胞中产生早期和晚期 rLEC。在目标 2 中，我们将确定 rLEC 对实验性淋巴水肿的治疗效果及其潜在机制这些治疗作用。在目标 3 中，我们将探讨 ETV2 是否能够直接将成纤维细胞重编程为功能性 LEC，诱导淋巴管形成，并改善淋巴水肿。我们预计结果所提出的实验将对新型细胞和基因疗法在治疗中的作用产生新的见解淋巴水肿。