tretinoin and Lymphedema

To determine the effect of 9-cis retinoic acid (9-cis RA) on postsurgical lymphedema.. 9-cis RA promotes lymphangiogenesis in vitro and in vivo and has promise as a therapeutic agent to limit the development of postsurgical lymphedema.. Lymphedema was induced in the right hind limb after a single fraction of 20 Gy radiation, popliteal lymphadenectomy, and lymphatic vessel ablation. Postoperatively, mice were randomly divided in to 2 groups that received daily intraperitoneal injections of either (1) an oil-based vehicle solution (control) or (2) 0.08 mg/kg of 9-cis RA dissolved in a vehicle solution. Outcome measures included paw thickness, lymphatic drainage, and lymphatic vessel density as measured by podoplanin immunohistochemistry and whole mount skin analysis.. Using our combined injury protocol, postsurgical lymphedema was observed 89% of the time. 9-cis RA-treated animals had less early postsurgical edema and significantly less paw lymphedema compared with vehicle-treated animals at all time-points (P < 0.001). 9-cis RA-treated animals had significantly faster lymphatic drainage as measured by indocyanine green clearance and increased lymphatic vessel density as measured by podoplanin immunohistochemistry (P < 0.001) and whole mount skin analysis (P < 0.05).. We have developed a highly reproducible model of secondary lymphedema and have demonstrated that 9-cis RA significantly prevents postsurgical lymphedema. Treatment with 9-cis RA is associated with increased lymphatic clearance and lymphangiogenesis. Because 9-cis RA (alitretinoin) is already approved for clinical use by the US Food and Drug Administration for other conditions, it has the potential to be repurposed as a preventative agent for postsurgical lymphedema in humans.

Topics: Alitretinoin; Animals; Antineoplastic Agents; Disease Models, Animal; Lymphangiogenesis; Lymphedema; Male; Mice; Mice, Transgenic; Postoperative Complications; Tretinoin

Lymphedema is mainly caused by lymphatic obstruction and manifested as tissue swelling, often in the arms and legs. Lymphedema is one of the most common post-surgical complications in breast cancer patients and presents a painful and disfiguring chronic illness that has few treatment options. Here, we evaluated the therapeutic potential of interleukin (IL)-8 in lymphatic regeneration independent of its pro-inflammatory activity. We found that IL-8 promoted proliferation, tube formation, and migration of lymphatic endothelial cells (LECs) without activating the VEGF signaling. Additionally, IL-8 suppressed the major cell cycle inhibitor CDKN1C/p57(KIP2) by downregulating its positive regulator PROX1, which is known as the master regulator of LEC-differentiation. Animal-based studies such as matrigel plug and cornea micropocket assays demonstrated potent efficacy of IL-8 in activating lymphangiogenesis in vivo. Moreover, we have generated a novel transgenic mouse model (K14-hIL8) that expresses human IL-8 in the skin and then crossed with lymphatic-specific fluorescent (Prox1-GFP) mouse. The resulting double transgenic mice showed that a stable expression of IL-8 could promote embryonic lymphangiogenesis. Moreover, an immunodeficient IL-8-expressing mouse line that was established by crossing K14-hIL8 mice with athymic nude mice displayed an enhanced tumor-associated lymphangiogenesis. Finally, when experimental lymphedema was introduced, K14-hIL8 mice showed an improved amelioration of lymphedema with an increased lymphatic regeneration. Together, we report that IL-8 can activate lymphangiogenesis in vitro and in vivo with a therapeutic efficacy in post-surgical lymphedema.

Topics: Animals; Cell Proliferation; Cells, Cultured; Cyclin-Dependent Kinase Inhibitor p57; Down-Regulation; Embryonic Development; Endothelial Cells; Homeodomain Proteins; Humans; Interleukin-8; Lymphangiogenesis; Lymphatic Vessels; Lymphedema; Mice; Mice, Transgenic; Neovascularization, Physiologic; Postoperative Complications; Receptors, Interleukin-8; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Regeneration; Tretinoin; Tumor Microenvironment; Tumor Necrosis Factor-alpha; Tumor Suppressor Proteins

Topics: Alitretinoin; Animals; Lymphangiogenesis; Lymphatic Vessels; Lymphedema; Regeneration; Tretinoin

The lymphatic system plays a key role in tissue fluid homeostasis and lymphatic dysfunction caused by genetic defects, or lymphatic vessel obstruction can cause lymphedema, disfiguring tissue swelling often associated with fibrosis and recurrent infections with no available cures to date. In this study, retinoic acids (RAs) were determined to be a potent therapeutic agent that is immediately applicable to reduce secondary lymphedema.. We report that RAs promote proliferation, migration, and tube formation of cultured lymphatic endothelial cells by activating fibroblast growth factor receptor signaling. Moreover, RAs control the expression of cell-cycle checkpoint regulators such as p27(Kip1), p57(Kip2), and the aurora kinases through both an Akt-mediated nongenomic action and a transcription-dependent genomic action that is mediated by Prox1, a master regulator of lymphatic development. Moreover, 9-cisRA was found to activate in vivo lymphangiogenesis in animals in mouse trachea, Matrigel plug, and cornea pocket assays. Finally, we demonstrate that 9-cisRA can provide a strong therapeutic efficacy in ameliorating experimental mouse tail lymphedema by enhancing lymphatic vessel regeneration.. These in vitro and animal studies demonstrate that 9-cisRA potently activates lymphangiogenesis and promotes lymphatic regeneration in an experimental lymphedema model, presenting it as a promising novel therapeutic agent to treat human lymphedema patients.

Topics: Alitretinoin; Animals; Aurora Kinases; Cell Movement; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p27; Cyclin-Dependent Kinase Inhibitor p57; Endothelial Cells; Fibroblast Growth Factors; Lymphangiogenesis; Lymphatic Vessels; Lymphedema; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Promoter Regions, Genetic; Protein Serine-Threonine Kinases; Regeneration; Tretinoin

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Azides; Cell Adhesion; Dendritic Cells; Deoxyribonucleases; Dogs; Edetic Acid; Endopeptidases; Epidermis; Heparin; Immunosuppressive Agents; Lidocaine; Lymph; Lymphedema; Lymphocytes; Neuraminidase; Sodium Azide; Tretinoin; Verapamil

To examine the mechanism of spontaneous attachment of afferent lymph lymphocytes to dendritic cells (veiled), we sampled and tested cells from skin lymph in 3 dogs with chronic lymphedema. There were 3.3 +/- 2.8% veiled cells in clusters with lymphocytes in lymph obtained directly from dilated dermal lymphatics by percutaneous puncture. The number of ex vivo clusters forming in the collected lymph samples increased as a function of time and was temperature dependent. The ability of veiled cells to bind lymphocytes was independent of divalent cations but reduced by xylocaine and retinoic acid. Among various steroids tested only methylprednisolone showed an inhibitory effect on cluster formation. Indomethacin and acetylsalicylic acid had no blocking activity on cell binding. Moreover, no effect was seen by cyclosporin A and azathioprine. However, FK 506 had a potent inhibitory effect on spontaneous cluster formation. This study suggests that cluster formation by skin lymph veiled cells and lymphocytes is a spontaneous process which can be controlled in vitro.

Topics: Animals; Anti-Inflammatory Agents; Dendritic Cells; Dogs; Edetic Acid; Immunosuppressive Agents; Lidocaine; Lymph; Lymphatic System; Lymphedema; Lymphocytes; Skin; Tretinoin; Verapamil