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The use of autologous cultured bone marrow-derived mesenchymal stem cells to treat venous ulcers: A pilot study A Grada, M Otero-Vin˜as, X Lin, T Yufit, P Carson and V Falanga Boston University School of Medicine, Boston, MA Background: Preclinical studies of mesenchymal stem cells in various animal models demonstrate accelerated wound healing through a variety of mechanisms. The aim of this small-scale study was to evaluate the effectiveness of autologous bone marrow-derived mesenchymal stem cells (BM-MSCs) in the treatment of patients with venous ulcers. Study design and method: A randomized, double- blind, placebo-controlled evaluation was selected to protect against bias. A total of 11 patients were randomly assigned to one of three groups: Group A (n¼4) received conventional standard therapy and control saline spray, group B (n¼3) received conventional standard therapy and fibrin spray, and group C (n¼4) received conventional standard therapy and BM-MSCs topically delivered using a fibrin spray. The total duration of the treatment was 24 weeks or earlier if the wound is completely closed. Patients were evaluated clinically at baseline and every 4 weeks up to week 24. The primary efficacy endpoint of the study is increased wound closure (cm/wk). Healing rates were calculated using the Gilman’s method. Data were analyzed by comparing the mean healing rates among treatment groups. Results: The average healing rate at week-4 for groups A, B, and C was 0.0006, -0.0522, and 0.1082 cm/wk, respectively. Patients received BM-MSCs treatment (Group C) showed higher mean healing rates and a significant reduction in wound size when compared to groups A and B at all evaluation time points. No adverse events were reported. Conclusions: This pilot study shows that topical application of autologous BM-MSCs may be an effective modality to promote healing in patients with difficult-to-heal wounds. However, larger studies are needed to confirm this finding.
Differential gene expression analysis of cutaneous lupus in mouse and human creates new insights into disease pathogenesis W Ko1, P Mande1, A Marshak-Rothstein1 and JE Harris2 1 UMass Medical School, Worcester, MA and 2 Department of Dermatology, University of Massachusetts Medical School, Worcester, MA Cutaneous lupus is a clinically heterogeneous disease, categorized into three main subtypes: discoid lupus (DLE), subacute cutaneous lupus (SCLE), and acute cutaneous lupus (ACLE). While these variants are clinically distinct, they appear to be similar on a pathologist’s histology slide. We utilized the Illumina HumanHT RNA microarray platform on formalin fixed, paraffin embedded biopsy sections of clinically diagnosed human cutaneous lupus (total n¼39; 6 DLE, 12 SCLE, 7 ACLE, 14 control) and analyzed the data using bioinformatics tools, R and Bioconductor. The data include relative expression levels for many transcripts in the human genome (n¼47304 probes). We identified several immune markers that were over- or under- expressed in all three disease conditions relative to control. For example, CXCL10 was increased 46.2 fold in DLE (p¼6.48e-11), 32.8 fold in SCLE (p¼2.15e-11), and 45.8 fold in ACLE (p¼1.24e-09). The preponderance of markers common to all disease subtypes suggests that lupus variants are indeed transcriptionally more similar than distinct, in accordance with histological examination. We also identified a few key differences among the disease states, such as IFN-e, which was relatively specific for DLE. We corroborated our findings in a novel mouse model of cutaneous lupus characterized by a prominent interface dermatitis, IgG deposition at the dermal-epidermal junction, mucin deposition, and extensive keratinocyte death. In one application, FAS Ligand was identified in the human data with increases of 3.28 fold in DLE (p¼6.11e-06), 3.08 fold in SCLE (p¼1.49e-06), and 3.07 fold in SLE (p¼8.63e05). The mouse model revealed that FasL knockout hosts were protected from disease, supporting an important role for FasL in the pathogenesis of cutaneous lupus. We anticipate that our human microarray analysis, combined with murine studies, will indicate new avenues for translational research.
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IL-22 promotes early aggressive behavior of cutaneous squamous cell carcinoma (SCC); A murine xenograft model M Abikhair1, N Roudiani1, A Santana1, J Chen2, D Felsen2 and J Carucci1 1 Dept of Derm, NYU Langone Med Ctr, New York, NY and 2 Inst for Ped Urol, Weill Cornell Medicine, New York, NY Immune-suppressed organ transplant recipients (OTRs) can develop aggressive SCC with extensive body surface area involvement and early metastasis. There are no curative systemic treatment options currently available. We have previously shown increased risk for catastrophic SCC in our OTR cohort via Cyclosporin A (CSA) mediated induction of IL-22 (Abikhair et al., JCI Insight, 2016; Zhang et al., PLoS ONE, 2013). In this study, we examined the link between IL-22 treatment, downstream signaling and aggressive SCC behavior both in vitro and in a murine xenograft model. We treated serum starved A431 SCC cells with IL-22 and found increased pSTAT3 activation within 15 minutes of treatment and significantly increased cell counts by 1 hour (1.25 fold, p < 0.05). A431 cells were injected into nude mice intradermally and mice were randomized to four treatment groups at day 7; IL-22 4ug daily, 5 days per week (n¼9), CSA 20mg/kg daily (n¼9), IL-22 plus CSA (n¼7) or vehicle (n¼9). Tumor size and volume were measured, and any ulceration documented. By the 5th day of treatment, IL-22 treated mice had significantly increased tumor growth from baseline compared to control (1.8 fold increase, p < 0.01). This growth difference decreased over the course of the experiment suggesting greatest effect at an early tumor stage. CSA treated mice had significantly greater tumor growth compared to control following 12 days of treatment. The combined treatment group showed earlier and a greater extent of tumor ulceration compared to vehicle. IL-22 hastens early aggressive behavior in SCC in vivo. This suggests there is a role for targeting the IL-22 axis in the early stage of SCC development, which may reduce the progression of aggressive SCC in immunosuppressed OTRs.
The role of MICA expression in acquired idiopathic generalized anhidrosis H Tanizaki1, S Moriwaki1 and K Kabashima2 1 Department of Dermatology, Osaka Medical College, Osaka, Japan and 2 Department of Dermatology, Kyoto Univ., Kyoto, Japan Acquired idiopathic generalized anhidrosis (AIGA) is characterized by generalized absence of sweating without other autonomic and neurologic dysfunction. There have been a considerable number of reports demonstrating that T lymphocytes infiltrate around eccrine glands and autoimmune mechanism is considered to play an important role in the development of AIGA. However, the pathogenesis of AIGA unknown. Normally, natural killer (NK) cells attack cells with absent/low MHC class I expression. However, because few NK cells are found around eccrine glands, we asked how eccrine glands escape from NK cell attack. Regarding the above, we performed histopathological analysis in anhidrotic, hypohidrotic, and normal sweating area. The eccrine glands and ducts on anhidrotic and hypohidrotic area were atrophy and surrounded by infiltrates of CD4-positive and CD8-positive lymphocytes. A considerable number of CD56-positive cells, Foxp3-positive cells, NKG2D-positive cells were observed around the eccrine glands. Major histocompatibility complex class I chainrelated A (MICA)-positive cells and decrease in expression of cholinergic receptors M3 were observed in anhidrotic sweating eccrine glands. These results indicate that MICA may evoke immune responses by interacting with NKG2D receptor on the surface of NK cells and CD8positive T cells and engagement of NKG2D by MICA triggers NK cells and co-stimulates some gd T cells and antigen-specific CD8 ab T cells. Thus, The MICA-NKG2D interaction may explain the possible pathogenic role in AIGA and we have to take into account in AIGA pathogenesis and its management.
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Analysis of sirolimus treatment response in lymphatic malformations with and without PIK3CA mutation MN Pyles1, JH Tu1, W Donahue2, E Tafoya1, Z Zinn1, A Marqueling1, MR Jeng3 and J Teng1 1 Stanford Dermatology, Stanford, CA, 2 Department of Pathology, University of Utah, Salt Lake City, UT and 3 Stanford Pediatric Hematology, Stanford, CA Sirolimus has been shown to be a highly effective medical therapy for lymphatic malformations (LM). Recent studies have demonstrated that many patients with lymphatic or venolymphatic malformations (VLM) have associated PIK3CA mutations. Our goal was to understand whether the presence of PIK3CA mutations among patients with LM or VLM would impact treatment response to Sirolimus. High-throughput sequencing was performed on archived tissue samples from 7 patients; blood samples were also obtained as controls. Volumetric analysis of pre-and post-treatment LM size on MRI was completed and adjusted for weight change. Five patients with confirmed PIK3CA mutations were treated with Sirolimus ranging 7e31 months, averaging 20.2 months. For these patients, decrease in LM size was positively correlated with Sirolimus treatment with an R2 value of 0.73, p-value ¼ 0.063. The average decrease in size of LM for this group was 36.5%. Genetic testing of two additional patients demonstrated no PIK3CA somatic mutation in their tissues. These patients were treated with Sirolimus for 9 and 35 months respectively. The average decrease in size of the LM was 29.4%. Genetic testing was not available for an additional 4 patients. Altogether, 11 patients demonstrated that decrease in LM size was positively correlated with Sirolimus treatment length with an R-value of 0.67, p-value ¼ 0.0018. Average change in LM size for all patients was -28.8%. As an mTOR inhibitor, Sirolimus is expected to be an effective agent in disorders in which the mTOR growth pathway is affected, which the PIK3CA mutation is known to affect. Aberrances in the mTOR pathway is likely causative for the LMs in this case but whether or not PIK3CA mutations impacts Sirolimus treatment outcomes still needs to be understood.
Analysis of thyroid function in 46 hospitalized patients with severe allergic dermatosis D Dong1, C Jin1, L Yang1 and Z Pan2 1 Department of Dermatology, Wuxi No.2People’s Hospital, Wuxi, China and 2 Department of Dermatology, Ren ji Hospital, School of Medicine, Shanghai JiaoTong University, Shanghai, China Objective: To study the thyroid function of 46 hospitalized patients with severe allergic dermatosis and its correlation with blood routine tests, high sensitivity C reactive protein and erythrocyte sedimentation rate. Methods: The test indexes of the thyroid function (including the TG-Ab and anti-TPOAb) about 46 hospitalized patients with acute urticaria (14/46), generalized eczema (13/46), allergic dermatitis (8/46), and drug eruption (11/46), were analyzed retrospectively. A correlation analysis was done between the thyroid function tests and blood routine tests, high sensitivity C reactive protein ,erythrocyte sedimentation rate. Results: The abnormal thyroid function results were found in 37 patients of 46 hospitalized patients (80.4%). The level of FT3, TT3 and TT4 was lower in contrast to the health control. The positive rate of the thyroid auto-antibody was higher than the health control (p < 0. 01). Further study showed the level of blood leukocyte, neutrophils, high sensitivity C reactive protein and erythrocyte sedimentation rate was higher than the health control respectively, while the level of hemoglobin was lower. In addition, the correlation analysis showed a positive correlation between hemoglobin and FT3, TT3 respectively, while a negative correlation was found between blood neutrophils and FT3, TT3 respectively. Conclusion: The abnormal thyroid function tests were common in patients with severe symptoms of allergic dermatosis. The main manifestation was the low T3 syndrome and the high thyroid autoantibody, which was correlated with the descend of hemoglobin and infection. These findings will help us to better understand severe allergic dermatosis and improve the treatment strategy.
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