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 淋巴水肿是由淋巴功能障碍引起的慢性肢体肿胀，影响全球2.5亿人。它 据估计，有5-10万美国人患有水肿，全世界有2.5亿人受到影响 继发性水肿最常发生在实体瘤的手术处理之后（例如，乳腺 癌症、黑素瘤）。腋窝淋巴结清扫术（ALND）用于局部晚期乳腺癌或 活检证实转移到腋窝淋巴结，并导致30%的患者术后出现水肿。 炎症引起的皮肤增厚、间质液潴留和皮下纤维脂肪沉积导致 进行性肢体增大淋巴水肿影响生活质量，并具有较高的健康成本负担。发病率 包括复发性蜂窝织炎、疼痛和肢体功能受损。 水肿的非手术治疗包括压迫疗法。手术治疗包括切除 (e.g皮肤/皮下切除术）和显微外科生理手术，包括血管化淋巴结 转移和淋巴静脉转流。目前的治疗方法可能会改善肢体大小和症状，但不能治愈 水肿由炎症表现引起的进行性肢体增大很难逆转。一 预防性手术策略称为立即淋巴重建（ILR）最近出现在 尝试减少水肿的频率。腋窝的传入神经纤维被破坏 在淋巴结切除过程中，通过显微外科手术将其与相邻静脉缝合。ILR减少了水肿 淋巴结清扫术后复发率为9%。然而，预防性ILR的广泛适用性是有限的， 需要专门的设备、技术专长、手术时间，并在选定的中心进行。 基于病毒载体的基因策略上调淋巴管生成的临床转化有限， 适用性病毒载体可在非预期位点引起全局淋巴管生成。组织纳米转染 已经开发了用于体内组织重编程的TNT技术。TNT有助于直接经皮 在快速（<100 ms）聚焦电场中使用用纳米通道制造的硅芯片进行基因递送。的 TNT用于基因递送的可行性已经被确立，并且在动物模型中的其他应用中得到验证。 我们提出了一种新颖的创新方法，在淋巴结转移时使用TNT进行非手术的局部基因递送。 损伤小鼠尾部水肿模型，以刺激淋巴管生成，防止淋巴功能障碍。 在目标1中，TNT将用于Prox 1的预防性基因递送，Prox 1是淋巴发育的主要调节因子 其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。在目标2中，我们将确定TNT递送基因的影响， 使用RNA-seq分析和细胞因子测定来研究炎症，以获得更精确的靶点。预防性 水肿显微外科（ILR）已经发展了水肿的外科治疗模式。这项探索性 关于脑水肿预防的建议将具有很高的转化意义和适应性。