title: Therapeutic effects of hyaluronidase on acquired lymphedema using a newly developed mouse limb model.
abstract: Acquired lymphedema is one of the most dreaded side effects of cancer treatment, such as surgical treatment or irradiation. However, due to the lack of appropriate animal models, there is no effective therapeutic method to cure acquired lymphedema. To develop a reproducible acquired lymphedema animal model, we devised a mouse hind limb model by removing a superficial inguinal lymph node, a popliteal lymph node, a deep inguinal lymph node, and the femoral lymphatic vessel. We measured the volume of lymphedematous leg and observed the change in level of hyaluronic acid (HA) and lymphangiogenic factors after injecting hyaluronidase. Our model showed the distinguishable swelling and the reliable symptoms compared to previously reported models. In the lymphedematous regions of our model, we confirmed that HA, a major component of extracellular matrix, accumulated to higher levels than in a normal mouse. This lymphedema volume was rapidly reduced by treating hyaluronidase. Following hyaluronidase injection, the lymphedematous region of our model resembled a normal hind limb. Our findings indicated that hyaluronidase promoted lymphangiogenesis on the lymphedematous limb. Based on hyaluronidase treatment in the lymphedematous region, this could potentially be a new therapeutic approach for acquired lymphedema mediated through the modification of the size of HA fragments. Impact statement In this manuscript, the essence of the work described in this manuscript involves the development of (1) a mouse limb model showing acquired lymphedema and (2) a potent therapeutic treatment using hyaluronidase to remedy acquired lymphedema in our model. In order to develop a reproducible acquired lymphedema animal model that reflects the most common symptoms experienced by lymphedema patients, we devised a mouse hind limb model by removing lymph nodes and lymphatics. Our model showed the distinguishable swelling and the reliable symptoms compared to previously reported models. In the lymphedematous regions of our model, we confirmed that hyaluronic acid (HA) accumulated to higher levels than in a normal mouse. This lymphedema volume was rapidly reduced by treating the lymphedematous leg with hyaluronidase, which also degraded high molecular weight HA to low molecular weight HA. Immunohistochemical analysis, quantitative real-time PCR analysis and lymphangioscintigraphy showed that hyaluronidase enhanced lymphangiogenesis in the lymphedematous limb.
authors.affiliation: 1 Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Kangsan
authors.initials: K
authors.lastname: Roh
authors.affiliation: 1 Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Sungrae
authors.initials: S
authors.lastname: Cho
authors.affiliation: 1 Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Jae-Hyun
authors.initials: JH
authors.lastname: Park
authors.affiliation: 2 Colorectal Cancer Branch, Division of Translational and Clinical Research I, Research Institute, National Cancer Center, Goyang 410-769, Republic of Korea.
authors.forename: Byong Chul
authors.initials: BC
authors.lastname: Yoo
authors.affiliation: 2 Colorectal Cancer Branch, Division of Translational and Clinical Research I, Research Institute, National Cancer Center, Goyang 410-769, Republic of Korea.
authors.forename: Won-Ki
authors.initials: WK
authors.lastname: Kim
authors.affiliation: 3 Molecular Imaging & Therapy Branch, National Cancer Center, Goyang 410-769, Republic of Korea.
authors.forename: Seok-Ki
authors.initials: SK
authors.lastname: Kim
authors.affiliation: 4 Department of Food Science and Biotechnology, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Kyewon
authors.initials: K
authors.lastname: Park
authors.affiliation: 5 Department of Oriental Medical Science, Graduate School of East-West Medicine, Kyunghee University, Secheondong, Yongin 446-701, Republic of Korea.
authors.forename: Hee
authors.initials: H
authors.lastname: Kang
authors.affiliation: 6 Natural Products Research Department, Gyeonggi Institute of Science & Technology, Suwon 443-270, Republic of Korea.
authors.forename: Jin-Mo
authors.initials: JM
authors.lastname: Ku
authors.affiliation: 7 U-Cell Clinic, Seoul 137-071, Republic of Korea.
authors.forename: Chang-Hwan
authors.initials: CH
authors.lastname: Yeom
authors.affiliation: 8 Department of Anatomy, School of Medicine, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Kyunghoon
authors.initials: K
authors.lastname: Lee
authors.affiliation: 1 Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Sukchan
authors.initials: S
authors.lastname: Lee
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D006614:Hindlimb; D006821:Hyaluronoglucosaminidase / Q000627:therapeutic use*; D008197:Lymph Node Excision; D008209:Lymphedema / Q000188:drug therapy* / Q000209:etiology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009369:Neoplasms / Q000150:complications
chemical_list: D006821:Hyaluronoglucosaminidase
keywords: Acquired lymphedema; hyaluronic acid; hyaluronidase and lymphangiogenesis