Prevention of Post-Surgical Lymphedema using Tissue Nanotransfection Technology

利用组织纳米转染技术预防术后淋巴水肿

• 批准号: 10810319
• 负责人: Aladdin Hasan Hassanein
• 金额: $ 38.36万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10810319
• 关键词: Affect; American; Anastomosis - action; Animal Model; Area; Axilla; Axillary Lymph Node Dissection; Axillary lymph node group; Biological Assay; Breast; Breast Cancer Treatment; Breast Cancer survivor; Breast biopsy; Bypass; Cellulitis; Chronic; Clinic; Consumption; Deposition; Disease; Dissection; Distal; Equipment; Etiology; Excision; Female; Frequencies; Gene Delivery; Genes; Genetic; Health Care Costs; Image; Impairment; Inflammation; Inflammatory; Inflammatory Response; Injury; Intercellular Fluid; Limb structure; Lymph; Lymph Node Dissections; Lymphangiogenesis; Lymphatic; Lymphatic Endothelial Cells; Lymphatic Endothelium; Lymphatic function; Lymphedema; Malignant Neoplasms; Mammaplasty; Mastectomy; Microsurgery; Modeling; Morbidity - disease rate; Mus; Neoplasm Metastasis; Operative Surgical Procedures; Pain; Patients; Persons; Physiological; Postoperative Period; Prevention; Procedures; Quality of life; Radiation; Recurrence; Research; Secondary to; Silicon; Site; Skin; Solid Neoplasm; Surgical Management; Swelling; Symptoms; Tail; Technical Expertise; Technology; Time; Tissue-Specific Gene Expression; Tissues; United States; United States National Institutes of Health; Up-Regulation; Vascularization; Veins; Viral Vector; advanced breast cancer; arm; cancer complication; clinical translation; compression therapy; cytokine; electric field; endothelial stem cell; iatrogenic injury; improved; in vivo; inflammatory marker; innovation; lymph nodes; lymphatic development; lymphatic dysfunction; malignant breast neoplasm; melanoma; mouse model; nanochannel; nanotransfection; novel; prevent; promoter; prophylactic; reconstruction; restoration; secondary lymphedema; soft tissue; subcutaneous; transcription factor; transcriptome sequencing

Project Summary Lymphedema is chronic limb swelling from lymphatic dysfunction which affects 250 million people worldwide. It is estimated that 5-10 million Americans have lymphedema, and 250 million people are affected worldwide Secondary lymphedema occurs most commonly following surgical management of solid tumors (e.g., breast cancer, melanoma). Axillary lymph node dissection (ALND) is performed for locally advanced breast cancer or biopsy-proven metastases to the axillary nodes and results in lymphedema in 30% of patients post-operatively. Skin thickening, interstitial fluid retention, and fibroadipose subcutaneous deposition from inflammation result in progressive limb enlargement. Lymphedema impacts quality of life and has a high health cost burden. Morbidity includes recurrent cellulitis, pain, and impaired extremity function. Non-surgical management of lymphedema includes compression therapy. Surgical treatment involves excisional (e.g, skin/subcutaneous resection) and microsurgical physiologic procedures including vascularized lymph node transfer and lymphovenous bypass. Current treatments may improve limb size and symptoms, but do not cure lymphedema. Progressive limb enlargement from the inflammatory manifestations is difficult to reverse. A preventative surgical strategy termed Immediate Lymphatic Reconstruction (ILR) has recently emerged in attempt to decrease the frequency of lymphedema. Afferent lymphatics in the axilla that have been disrupted during lymph node removal are microsurgically anastomosed to adjacent veins. ILR decreases the lymphedema occurrence to 9% after lymphadenectomy. However, broad applicability of preventative ILR is limited as it requires specialized equipment, technical expertise, prolongs operative time, and is performed in select centers. Viral vector-based gene strategies to upregulate lymphangiogeneis have had limited clinical translational applicability. Viral vectors can cause global lymphangiogenesis at unintended sites. Tissue nanotransfection technology (TNT) has been developed for in vivo tissue reprogramming. TNT facilitates direct, transcutaneous gene delivery using a silicon chip fabricated with nanochannels in a rapid (<100ms) focused electric field. The feasibility of TNT for gene delivery has been established and validated for other applications in animal models. We propose a novel, innovative approach to use TNT for non-surgical, focal gene delivery at the time of lymphatic injury to the murine tail model of lymphedema to stimulate lymphangiogenesis to prevent lymphatic dysfunction. In Aim 1, TNT will be used for prophylactic gene delivery of Prox1, a master regulator of lymphatic development which controls lymphatic endothelial progenitor cells and regulates lymphangiogenesis. Lymphatic function and lymphangiogenesis will be rigorously assessed. In Aim 2, we will determine the effects of TNT-delivered genes on inflammation using RNA-seq analysis and cytokine assays to lead to more precise targets. Prophylactic lymphedema microsurgery (ILR) has evolved the surgical treatment paradigm of lymphedema. This exploratory proposal on lymphedema prevention will have high translational significance and adaptability.

项目摘要 淋巴水肿是由于淋巴功能障碍导致的慢性肢体肿胀，全世界有2.5亿人受到影响。它 据估计，美国有500-1000万人患有淋巴水肿，全球有2.5亿人受到影响 继发性淋巴水肿最常发生在实体肿瘤(例如，乳腺)的外科治疗之后 癌症、黑色素瘤)。腋窝淋巴结清扫术(ALND)适用于局部晚期乳腺癌或 术后30%的患者经活检证实转移至腋窝淋巴结并导致淋巴水肿。 皮肤增厚、间质液体滞留和炎症导致的皮下纤维脂肪沉积 进行性肢体增大。淋巴水肿影响生活质量，并造成很高的医疗费用负担。发病率 包括复发性蜂窝组织炎、疼痛和肢体功能受损。 淋巴水肿的非手术治疗包括压迫治疗。手术治疗包括切除 (例如，皮肤/皮下切除)和显微外科生理手术，包括带血管的淋巴结 转流和淋巴静脉转流术。目前的治疗方法可能会改善肢体大小和症状，但不能治愈 淋巴浮肿。进行性肢体肥大从炎症性表现难以逆转。一个 最近出现了一种名为立即淋巴重建(ILR)的预防性手术策略 尝试减少淋巴水肿的频率。腋下的传入淋巴管已被破坏 在切除淋巴结的过程中，通过显微外科手术将其与邻近静脉吻合。ILR可减轻淋巴水肿 淋巴清扫术后发生率为9%。然而，预防性ILR的广泛适用性受到限制，因为它 需要专门的设备和技术专长，延长手术时间，并在选定的中心进行。 基于病毒载体的上调淋巴管生成的基因策略在临床上的应用有限 适用性。病毒载体可以在非预期位置引起全球淋巴管生成。组织纳米转基因 已经开发出用于体内组织重新编程的技术(TNT)。TNT促进了直接、经皮 在快速(100毫秒)聚焦电场中使用由纳米通道制成的硅芯片进行基因传递。这个 已经建立了TNT用于基因传递的可行性，并验证了TNT在动物模型中的其他应用。 我们提出了一种新颖的创新方法，在淋巴转移时使用TNT进行非手术的、局部的基因传递。 通过损伤小鼠尾部淋巴水肿模型，刺激淋巴管生成，预防淋巴功能障碍。 在目标1中，TNT将用于预防性地传递Prox1的基因，Prox1是淋巴发育的主要调节因子 它控制淋巴管内皮祖细胞并调节淋巴管生成。淋巴功能和 将严格评估淋巴管生成情况。在目标2中，我们将确定TNT传递的基因的影响 在炎症方面，使用RNA-seq分析和细胞因子分析以获得更精确的靶点。预防用药 淋巴水肿显微手术(ILR)发展了淋巴水肿的外科治疗模式。这种探索性的 关于预防淋巴水肿的建议将具有很高的翻译意义和适应性。