As research continues and our longing increases to find a cure for lymphedema, this study was sent to me by a membe of our Children with Lymphedema Yahoo Support Group.
I attempted to access the original site it came from in Cuba, but was unfortunately blocked, either by Google or by some Cuban governmental restriction. Never the less, I thought it important enough to post.
There is the possibility that this can be of tremendous value to us and I can only hope that researchers in the United States or other countries will move quickly into this new area of study.
As I can find further information, I will immediately update this page.
September 11, 2009
From EMORY Health Now Blog
Lymphedema, or swelling because of the impaired flow of lymph fluid, can occur as a consequence of cancer or cancer treatment. Chemotherapy can damage lymph ducts, and often surgeons remove lymph nodes that may be affected by cancer metastasis. Lymphedema can result in painful swelling, impaired mobility and changes in appearance.
Emory scientists, led by cardiologist and stem cell biologist Young-sup Yoon, have shown that they can isolate progenitor cells for the lining of lymph ducts. This finding could lead to doctors being able to regenerate and repair lymph ducts using a patient’s own cells. The results are described in a paper published recently in the journal Circulation.
The authors used the cell surface marker podoplanin as a handle for isolating the progenitor cells from bone marrow. Previous research has demonstrated that podoplanin is essential for the development of the lymphatic system. In the paper, the authors use several animal models to show that the progenitor cells could contribute to the formation of new lymph ducts, both by becoming part of the lymph ducts and by stimulating the growth of nearby cells. “This lymphatic vessel–forming capability can be used for the treatment of lymphedema or chronic unhealed wounds,” Yoon says.
The authors also show that mice with tumors show an increase in the number of this type of circulating progenitor cells. This suggests that tumors send out signals that encourage lymph duct growth – a parallel to the well-known ability of tumors to drive growth of blood vessels nearby. Yoon says the presence of these cells could be a marker for tumor growth and metastasis. Because tumors often metastasize along lymph ducts and into lymph nodes, studying this type of cells could lead to new targets for blocking tumor metastasis. A recent review in the journal Genes & Development summarizes additional functions of the lymphatic system in fat metabolism, obesity, inflammation, and the regulation of salt storage in hypertension.
Editor's note: Last year I was able to get to know Dr. Yoon at our Georgia Lymphedema Awareness program. I must say that I was very impressed by both his knowledge and his compassion and drive to find a cure for those of us with lymphedema. Pat O'Connor
Yan A, Avraham T, Zampell JC, Haviv YS, Weitman E, Mehrara BJ.
The Division of Plastic & Reconstructive Surgery, Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA; 1275 York Avenue, Room MRI 1005, New York, NY 10065, USA.
Keywords: adipose-derived stem cells, antilymphangiogenic, lymphangiogenesis, TGF-β, tissue engineering, VEGF-C
Aims: Recent studies have demonstrated that augmentation of lymphangiogenesis and tissue engineering hold promise as a treatment for lymphedema. The purpose of this study was to determine whether adipose-derived stem cells (ASCs) can be used in lymphatic tissue-engineering by altering the balance between pro- and anti-lymphangiogenic cytokines.
Materials & methods ASCs were harvested and cultured in media with or without recombinant VEGF-C for 48 h. ASCs were then implanted in mice using Matrigel plugs. Additional groups of animals were implanted with ASCs transfected with a dominant-negative TGF-β1 receptor-II adenovirus with or without VEGF-C stimulation, since TGF-β1 has been shown to have potent antilymphangiogenic effects. Lymphangiogenesis, lymphatic differentiation and cellular proliferation were assessed.
Results: Stimulation of ASCs with VEGF-C in vitro significantly increased expression of VEGF-A, VEGF-C and Prox-1. ASCs stimulated with VEGF-C prior to implantation induced a significant (threefold increase) lymphangiogenic response as compared with control groups (unstimulated ASCs or empty Matrigel plugs; p < 0.01). This effect was significantly potentiated when TGF-β1 signaling was inhibited using the dominant-negative TGF-β1 receptor-II virus (4.5-fold increase; p < 0.01). Stimulation of ASCs with VEGF-C resulted in a marked increase in the number of donor ASCs (twofold; p < 0.01) and increased the number of proliferating cells (sevenfold; p < 0.01) surrounding the Matrigel. ASCs stimulated with VEGF-C expressed podoplanin, a lymphangiogenic cell marker, whereas unstimulated cells did not.
Conclusion: Short-term stimulation of ASCs with VEGF-C results in increased expression of VEGF-A, VEGF-C and Prox-1 in vitro and is associated with a marked increase lymphangiogenic response after in vivo implantation. This lymphangiogenic response is significantly potentiated by blocking TGF-β1 function. Furthermore, stimulation of ASCs with VEGF-C markedly increases cellular proliferation and cellular survival after in vivo implantation and stimulated cells express podoplanin, a lymphangiogenic cell marker.
Editor's note: VEGF-C is critical to trigger formation of new lymphatics. Therefore this could prove to be a major step in this promising lymphedema treatment.
Maldonado GE, Pérez CA, Covarrubias EE, Cabriales SA, Leyva LA, Pérez JC, Almaguer DG.
Hospital Universitario 'Dr José Eleuterio González', Universidad Autónoma de Nuevo León, México. firstname.lastname@example.org
Keywords: autologous; lymphedema; mastectomy; stem cells
BACKGROUND AIMS. Lymphedema is a common complication with breast cancer treatment that does not have a definite cure. Our objective was to determine the efficacy of autologous stem cells (ASC) in the treatment of lymphedema secondary to mastectomy and axillary lymphadenectomy in comparison with traditional decongestive treatment with compression sleeves.
METHODS. A prospective study including 20 women with lymphedema secondary to breast cancer surgery with axillary lymphadenectomy was conducted. Women were assigned at random to one of two groups. One group of 10 women was injected with ASC in the affected arm, whereas the other 10 women comprised the control group and received traditional compression sleeve therapy (CST). The follow-up for both groups was 12 weeks. Pain, sensitivity and mobility were assessed before and after therapy.
RESULTS. There was improvement in the volume of lymphedema in both groups, with no significant difference. In the ASC group there was an overall volume reduction during the follow-up, whereas in the CST group lymphedema recurred after the compression sleeve was removed.
CONCLUSIONS. Our findings suggest that ASC injection for patients with lymphedema can be an effective treatment. It reduces arm volume and associated co-morbidities of pain and decreased sensitivity. Traditional CST was also effective for lymphedema reduction, but it was dependent on continuous use of the treatment.
Razmkhah M, Jaberipour M, Erfani N, Habibagahi M, Talei AR, Ghaderi A.
Shiraz Institute for Cancer Research, Shiraz University of Medical Sciences, Shiraz, Iran.
Keywords: Adipose derived stem cells (ASCs); Breast cancer; Tumor microenvironment; IL-10; IL-4; TGF-β1
Immunomodulatory function of bone marrow derived mesenchymal stem cells in cancer has recently been investigated. But the resident mesenchymal stem cells as whole in cancer and in the breast cancer tissue have not been studied well. In the present work we isolated adipose derived stem cells (ASCs) from breast cancer and normal breast tissues to investigate the expressions of IL-4, IL-10 and transforming growth factor (TGF)-β1 in ASCs and to see if ASCs isolated from patients can modulate the regulatory molecules on peripheral blood lymphocytes.
Our results showed that IL-10 and TGF-β1 have significantly higher mRNA expressions in ASCs isolated from breast cancer patients than those from normal individuals (P value <0.05). The culture supernatant of ASCs isolated from breast cancer patients with pathological stage III induced upregulation of the mRNA expression levels of IL-4, TGF-β1, IL-10, CCR4 and CD25 in PBLs.
In addition, the percentage of CD4+CD25(high)Foxp3(+) T regulatory cells was increased in vitro. When the same culture supernatant was added to ASCs isolated from normal subjects augmentation of the mRNA expressions of IL-4, IL-10, IL-8, MMP2, VEGF and SDF-1 in normal ASCs was also observed. These data collectively conclude that resident ASCs in breast cancer tissue may have crucial roles in breast tumor growth and progression by inducing regulatory molecules and promoting anti-inflammatory reaction within the tumor microenvironment.
Further investigation is required to see if the immune suppression induced by ASCs is an independent property from tumor cells or ASCs gain their immunosuppressive potential from malignant cells.
Hwang JH, Kim IG, Lee JY, Piao S, Lee DS, Lee TS, Ra JC, Lee JY.
Department of Physical and Rehabilitation Medicine, Sungkyunkwan University School of Medicine, Samsung Medical Center, Seoul, Republic of Korea.
Keywords: Lymphangiogenesis; Stem cell; Hydrogel; VEGF-C
Lymphedema is a manifestation of lymphatic system insufficiency. It arises from primary lymphatic dysplasia or secondary obliteration after lymph node dissection or irradiation. Although improvement of swelling can be achieved by comprehensive non-operative therapy, treatment of this condition requires lifelong care and good compliance. Recently molecular-based treatments using VEGF-C have been investigated by several researchers. We designed the present study to determine whether the therapeutic efficacy of implanted human adipose-derived stem cells (hADSCs) could be improved by applying a gelatin hydrogel containing VEGF-C (VEGF-C hydrogel) to the site of tissue injury in a lymphedema mouse model. Four weeks after the operation, we evaluated edema and determined lymphatic vessel density at various post-operative time points. Mice treated with hADSCs and VEGF-C hydrogel showed a significantly decreased dermal edema depth compared to the groups of mice that received hADSCs only or VEGF-C hydrogel only.
Immunohistochemical analysis also revealed that the hADSC/VEGF-C hydrogel group showed significantly greater lymphatic vessel regeneration than all the other groups. hADSCs were detected in the implantation sites of all mice in the hADSC/VEGF-C group, and exhibited a lymphatic endothelial differentiation phenotype as determined by co-staining PKH-labeled hADSCs for the lymphatic marker LYVE-1. Our results suggest that co-administration of hADSCs and VEGF-C hydrogel has a substantial positive effect on lymphangiogenesis.
September 10, 2009
Goicoechea Díaz P, Artaza Sanz HM, Blanco Díaz A, García Pelegrin S, Atencio Sariol E, Hernández Ramírez P1, González Suárez T1, Matamoros Martínez de Pinillos MA1, Socarrás Ferrer BB1, del Valle Pérez L1, Peña Quian Y2, Pintado Perera A2.
Hospital Clínico Quirúrgico Docente “Enrique Cabrera”. La Habana, 1Instituto de Hematología e Inmunología. La Habana, 2Centro de Investigaciones Clínicas. La Habana. Cuba. email@example.com
Lymphedema is a chronic disease characterized by abnormal accumulation of lymph due to a failure of the lymphatic system, which can affect the upper limbs, lower and external genitalia. The lymphedema are classified as primary if they have no identifiable cause defects if present from birth, early or late depending on the age at which they appear and side, those with an identifiable cause, including those appearing post -lymphangitis, post radiation, for filariasis.
In this study, we evaluated in patients with lower limb lymphedema in the efficacy of autologous stem cell implants derived from bone marrow and mobilized peripheral blood by stimulation with granulocyte colony stimulating factor.
We treated 6 patients, 3 men and 3 women, mean age 48 years, all with lymphedema in both lower limbs by recurrent lymphangitis. We performed the implantation of stem cells in the most affected leg. The injections were made at a distance of 3 cm between them and a volume of 1 ml per puncture, varying the total volume according to the intensity and extent of lymphedema. In all cases there has been a progressive decrease in diameter of the injected limb and has been an improvement in the evolutionary lymphography performed.
We believe this approach is an alternative method to treat this disease, which over the years has not benefited from the treatment methods normally used.
Kreuger J, Nilsson I, Kerjaschki D, Petrova T, Alitalo K, Claesson-Welsh L.
Department of Genetics and Pathology, Uppsala University, Sweden. firstname.lastname@example.org
Key Words: embryoid body; lymphangiogenesis; LYVE-1; Prox1; VEGF-C; VEGF receptor-3
OBJECTIVE: The purpose of this study was to establish a model system for lymph vessel development based on directed differentiation of murine embryonic stem cells.
METHODS AND RESULTS: Stem cells were aggregated to form embryoid bodies, and subsequently cultured in 3-dimensional collagen matrix for up to 18 days. Treatment with vascular endothelial growth factor (VEGF)-C and VEGF-A individually enhanced formation of lymphatic vessel structures, although combined treatment with VEGF-C and VEGF-A was most potent and gave rise to a network of LYVE-1, podoplanin, Prox1, and VEGF receptor-3 positive lymphatic vessel structures running parallel to and apparently emanating from, capillaries. In contrast, fibroblast growth factor-2, hepatocyte growth factor, or hypoxia had little or no effect on the development of the early lymphatics.
Further, cells of hematopoietic origin were shown to express lymphatic markers. In summary, different subpopulations of lymphatic endothelial cells were identified on the basis of differential expression of several lymphatic and blood vessel markers, indicating vascular heterogeneity.
CONCLUSIONS: We conclude that the present model closely mimics the early steps of lymph vessel development in mouse embryos.
Full Text Article American Heart Association
Kono T, Kubo H, Shimazu C, Ueda Y, Takahashi M, Yanagi K, Fujita N, Tsuruo T, Wada H, Yamashita JK.
Molecular and Cancer Research Unit, HMRO and Department of Thoracic Surgery, Graduate School of Medicine, Kyoto University, Japan; Laboratory of Stem Cell Differentiation, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Japan; Institute of Molecular and Cellular Biosciences, The University of Tokyo, Japan; and PRESTO, Japan Science and Technology Agency, Japan.
Correspondence to Hajime Kubo, Molecular and Cancer Research Unit, HMRO, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan. E-mail email@example.com; or Jun K. Yamashita, Laboratory of Stem Cell Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Shogoin-Kawahara-cho53, Sakyo-ku, Kyoto, 606-8507, Japan. E-mail firstname.lastname@example.org
Key Words: lymphatic endothelial cells; embryonic stem cells; prox1; VEGF-C; VEGFR3
OBJECTIVE: The discovery of vascular endothelial growth factor C (VEGF-C) and VEGF receptor-3 (VEGFR-3) has started to provide an understanding of the molecular mechanisms of lymphangiogenesis. The homeobox gene prox1 has been proven to specify lymphatic endothelial cells (ECs) from blood ECs. We investigated the process of lymphatic EC (LEC) differentiation using embryonic stem (ES) cells.
METHODS AND RESULTS: VEGFR-2(+) cells derived from ES cells differentiated into LECs at day 3 on OP9 stromal cells defined by the expression of prox1, VEGFR-3, and another lymphatic marker podoplanin. VEGFR-2(+) cells gave rise to LYVE-1(+) embryonic ECs, which were negative for prox1 on day 1 but turned to prox1(+) LECs by day 3. VEGFR-3-Fc or Tie2-Fc, sequestering VEGF-C or angiopoietin1 (Ang1), suppressed colony formation of LECs on OP9 cells. However, addition of VEGF-C and Ang1 in combination with VEGF to the culture of VEGFR-2(+) cells on collagen-coated dishes failed to induce LECs. LEC-inducing activity of OP9 cells was fully reproduced on paraformaldehyde-fixed OP9 cells with the conditioned medium.
CONCLUSIONS: We succeeded in differentiating LECs from ES cells and revealed the requirements of VEGF-C, Ang1, and other unknown factors for LEC differentiation.
PMID: 16690875 [PubMed - as supplied by publisher]
Full Text Article American Heart Association
Hirashima M, Ogawa M, Nishikawa S, Matsumura K, Kawasaki K, Shibuya M, Nishikawa S. Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Japan.
Keywords: VEGF; VEGFR2; CD31; CD34; TIE2/TEK; Flt-1; mesodermal cells; Monoclonal antibodies; MoAbs; VE-cadherin; VECD1
Vascular endothelial growth factor (VEGF) is a major growth factor for developing endothelial cells (ECs). Embryonic lethality due to haploinsufficiency of VEGF in the mouse highlighted the strict dose dependency of VEGF on embryonic vascular development. Here we investigated the dose-dependent effects of VEGF on the differentiation of ES cell-derived fetal liver kinase 1 (Flk-1)/VEGF receptor 2(+) (VEGFR2(+)) mesodermal cells into ECs on type IV collagen under a chemically defined serum-free condition. These cells could grow even in the absence of VEGF, but differentiated mostly into mural cells positive for alpha-smooth muscle actin.
VEGF supported in a dose-dependent manner the differentiation into ECs defined by the expression of VE-cadherin, platelet-endothelial cell adhesion molecule 1 (PECAM-1)/ CD31, CD34, and TIE2/TEK. VEGF requirement was greater at late than at early phase of culture during EC development, whereas response of VEGFR2(+) cells to VEGF-E, which is a virus-derived ligand for VEGFR2 but not for Flt-1/VEGFR1, was not dose sensitive even at late phase of culture. Delayed expression of VEGFR1 correlated with increased dose dependency of VEGF.
These results suggested that greater requirement of VEGF in the maintenance than induction of ECs was due to the activity of VEGFR1 sequestering VEGF from VEGFR2 signal. The chemically defined serum-free culture system described here provides a new tool for assessing different factors for the proliferation and differentiation of VEGFR2(+) mesodermal cells.
Full text article Journal of the American Society of Hematology
Keywords: Breast cancer metastasis; Cancer invasion; Carboxylesterase; CPT-11;Cytokines; Interleukin-6; Neural stem cells; siRNA;Stem cell tropism; Stem cell-mediated therapy; Gene therapy
Keywords: Adipose stem cells; Stem cell therapy; Good manufacturing practice; Defined serum-free culturing conditions; Flow cytometry
Key words Adult stem cells; Adipose-derived stem cells/stromal cells; Adipose tissue; Regenerative medicine
Keywords: Monocytes; Lymphatic phenotypes; VEGF-C; LYVE-1; Prox1; Podoplanin; endothelial markers; fibronectin (FN)
Keywords: Adipose-derived stromal cells; Skeletal tissue engineering; Tissue regeneration; Multipotent stromal cells; Adipogenic differentiation; Subcutaneous fat depots
Keywords: disc degeneration; gene therapy; nucleus pulposus cell; Sox-9;stem cells; type II collagen
Keywords: adipose tissue; adipose tissue-derived stem cells; angiogenesis; bone marrow; cell therapy; myocardial regeneration; myocardial repair; myogenic differentiation; stem cells; stromal cells; tissue engineering
Key Words: lymphangiogenesis; Lyve-1; mesenchymal stem cell; Prox-1; VEGF; Prox-1; podoplanin; VEGF receptor-2; VEGF receptor-3