RESUMEN
Epidermal transplantation is a common and widely used surgical technique in clinical medicine. Derivatives of embryonic stem cells have the potential to serve as a source of transplantable cells. However, allograft rejection is one of the main challenges. To investigate the immunogenicity of keratinocytes derived from human embryonic stem cells (ESKCs), we conducted a series of in vivo and in vitro experiments. The results showed that ESKCs have low HLA molecule expression, limited antigen presentation capabilities, and a weak ability to stimulate the proliferation and secretion of inflammatory factors in allogeneic PBMCs in vitro. In humanized immune mouse models, ESKCs elicited weak transplant rejection responses in the host. Overall, we found that ESKCs have low immunogenicity and may have potential applications in the field of regenerative medicine.
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Células Madre Embrionarias Humanas , Queratinocitos , Humanos , Queratinocitos/inmunología , Queratinocitos/metabolismo , Queratinocitos/citología , Células Madre Embrionarias Humanas/citología , Células Madre Embrionarias Humanas/inmunología , Células Madre Embrionarias Humanas/metabolismo , Animales , Ratones , Proliferación Celular , Rechazo de Injerto/inmunología , Antígenos HLA/inmunología , Antígenos HLA/metabolismoRESUMEN
BACKGROUND: Surgical intervention is the main therapy for refractory vitiligo. We developed a modified autologous cultured epithelial grafting (ACEG) technique for vitiligo treatment. Between January 2015 and June 2019, a total of 726 patients with vitiligo underwent ACEG in China, with patient characteristics and clinical factors being meticulously documented. Using a generalized linear mixed model, we were able to assess the association between these characteristics and the repigmentation rate. RESULTS: ACEG demonstrated a total efficacy rate of 82.81% (1754/2118) in treating 726 patients, with a higher repigmentation rate of 64.87% compared to conventional surgery at 52.69%. Notably, ACEG showed a better response in treating segmental vitiligo, lesions on lower limbs, ageâ ≤â 18, and stable periodâ >â 3 years. A keratinocyte:melanocyte ratio below 25 was found to be advantageous too. Single-cell RNA sequencing analysis revealed an increase in melanocyte count and 2 subclusters of keratinocytes after ACEG, which remained higher in repigmented sites even after 1 year. CONCLUSIONS: ACEG is a promising therapy for refractory vitiligo. Patient age, clinical type, lesion site, and stability before surgery influence repigmentation in ACEG. The mechanism of repigmentation after ACEG treatment is likely not confined to the restoration of melanocyte populations. It may also involve an increase in the number of keratinocytes that support melanocyte function within the affected area. These keratinocytes may aid the post-transplant survival and function of melanocytes by secreting cytokines and extracellular matrix components. TRIAL REGISTRATION: registered with Chictr.org.cn (ChiCTR2100051405).
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Trasplante Autólogo , Vitíligo , Humanos , Vitíligo/terapia , Masculino , Femenino , Estudios Retrospectivos , Trasplante Autólogo/métodos , Adulto , Adolescente , Adulto Joven , Persona de Mediana Edad , Melanocitos/trasplante , Niño , Queratinocitos/trasplante , Células Cultivadas , EpitelioRESUMEN
Autologous cultured epithelium grafting (ACEG) presents a promising treatment for refractory vitiligo, yet concerns regarding infections and immunological reactions hinder its surgical use due to serum and feeder dependencies. Addressing this, we culture autologous epithelium under serum- and feeder-free (SFF) conditions, comparing its safety and efficacy with serum- and feeder-dependent (SFD) conditions in stable vitiligo patients, and we discover no significant differences in repigmentation between the SFF and SFD grafts. Single-cell RNA transcriptomics on SFF- and SFD-cultured epithelium alongside healthy skin reveal increased populations of LAMB3+ basal keratinocytes and ZNF90+ fibroblasts in the SFF sheets. Functional analyses showcase active cellular metabolism in LAMB3+ basal keratinocytes, vital in extracellular matrix homeostasis, while ZNF90+ fibroblasts demonstrate increased differentiation, essential in collagen formation for cell adhesion. Importantly, these cell populations in SFF sheets exhibit enhanced interactions with melanocytes compared to SFD sheets. Further, knockdown experiments of LAMB3 in keratinocytes and ZNF90 in fibroblasts lead to a downregulation in melanocyte ligand-receptor-related genes. Overall, SFF sheets demonstrate comparable efficacy to SFD sheets, offering superior safety. LAMB3+ basal keratinocytes and ZNF90+ fibroblasts act as potential drivers behind repigmentation in ACEG under SFF conditions. This study provides translational insights into ACEG repigmentation and potential therapeutic targets for vitiligo.
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Vitíligo , Humanos , Vitíligo/terapia , Epitelio , Queratinocitos , Piel , FibroblastosRESUMEN
The study investigated the effectiveness of EDN1 and EDN3 cytokines in the differentiation of melanocytes from hESCs. The findings showed that 100 nM EDN1 was more effective in promoting hESC to CD117+/TYR+ melanoblasts compared to 100 nM EDN3. Additionally, maintaining melanoblasts is beneficial for preserving the ability to proliferate. The study found that 10 nM EDN1 helped maintain the proliferation of melanoblasts without over maturing them into melanocytes in the late stage of differentiation. Thus, using 100 nM EDN1 in the initial stage and 10 nM EDN1 in the late stage proved to be an efficient and cost-effective method for obtaining hESC-derived melanocytes. The preliminary results suggest that EDN1 promotes melanoblast formation during the initial differentiation stage through its binding to both the EDNRB receptor and EDNRA receptor. This study provides a valuable tool for studying the development of human melanocytes and modelling the biology of disease.
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Endotelina-1 , Células Madre Embrionarias Humanas , Humanos , Endotelina-1/metabolismo , Melanocitos/metabolismo , Diferenciación CelularRESUMEN
Although surgical interventions have become optional for refractory vitiligo, grafting related injuries is inevitable. Embryonic stem cell (ESC) derivatives can be used in transplantation to address this issue, but the immune rejection due to allogeneic transplantation is of great concern. To investigate the immunogenicity of ESC derived melanocytes (ES-MC), we established a co-culture system of ES-MC and allogeneic PBMC. The results showed that ES-MC were similar to human primary melanocytes, with low expression of immune related molecules, and limited capability of stimulating allogeneic lymphocytes in vitro. Taken together, our findings confirm that ES-MC are of limited immunogenicity, providing new insights into the application of ES-MC in the regenerative medicine such as treating vitiligo.
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Células Madre Embrionarias Humanas/inmunología , Melanocitos/inmunología , Diferenciación Celular/inmunología , Células Cultivadas , Técnicas de Cocultivo , Células Madre Embrionarias Humanas/citología , Humanos , Melanocitos/citología , Medicina RegenerativaRESUMEN
PURPOSE: Our study was to investigate 18F-FLT PET/CT imaging monitor the early response of CDK4/6 inhibitor therapy in triple negative breast cancer (TNBC). METHODS: MDA-MB-231 and MDA-MB-468 cell lines and corresponding subcutaneous tumor models in CB17-SCID mice were used. Cell viability assay, cell-cycle analysis, and western blotting were performed in vitro experiments. 18F-FLT PET/CT imaging was performed and the value of tumor/muscle (T/M) of mice was measured before and 1-3 days after treatment in vivo experiments. Then, the tumor volume was recorded every day for 15 days. RESULTS: In the presence of Palbociclib (CDK4/6 inhibitor), the results of in vitro experiments showed that protein pRB and E2F levels were significantly down-regulated in MDA-MB-231 cells leading to G0/G1 arrest with consumption in S phase compared with MDA-MB-468 cells. In PET/CT imaging, the 18F-FLT T/M ratio of treatment group was a significant and sustained reduction from 1 to 3 days (all p < 0.05) compared with control group in MDA-MB-231 section. However, there was no significant difference between treatment and control groups in MDA-MB-468 section. Compared with the control group, the tumor volume of the treatment group was significantly reduced from the 11th day in MDA-MB-231 section, but not in MDA-MB-468 section until 15 days. CONCLUSION: 18F-FLT PET/CT imaging can immediately and effectively monitor the early treatment response of CDK4/6 inhibitors in TNBC.
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Tomografía Computarizada por Tomografía de Emisión de Positrones , Neoplasias de la Mama Triple Negativas/diagnóstico por imagen , Animales , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Supervivencia Celular , Quinasa 4 Dependiente de la Ciclina , Femenino , Ratones , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
OBJECTIVE: To compare the predictive value of European Organization for Research and Treatment of Cancer (EORTC) criteria and PET Response Criteria in Solid Tumors (PERCIST), for the pathological response and prognosis of patients with breast cancer receiving neoadjuvant chemotherapy (NAC). METHODS: Consecutive PET/computed tomography scans in 128 operable female patients at baseline and after two courses of NAC were performed. Patients were categorized by complete metabolic response (CMR) and non-CMR groups using 2 PET criteria. CMR and non-CMR were used to predict pathological complete response (pCR) by diagnostic test evaluation, and to predict progression-free survival (PFS) using Kaplan-Meier plots and Cox proportional hazards regression. RESULTS: Ninety-two patients were finally analyzed. The sensitivity, specificity, and accuracy for pCR prediction were 69.7, 76.3, and 73.9% with EORTC criteria, and 69.7, 77.9, and 75.0% with PERCIST, respectively. Peak standardized uptake value normalized to lean body mass (SULpeak), maximum standardized uptake value (SUVmax), total lesion glycolysis (TLG), and metabolic tumor volume (MTV) were pCR response with accuracy of 70.7, 60.0, 75.0, and 71.4%, respectively. CMR by the EORTC (P = 0.021) and PERCIST (P = 0.007) was significantly related to a longer PFS. The univariate and multivariate analysis suggested that CMR by PERCIST was an independent predictor of recurrence (P = 0.008). CONCLUSION: EORTC criteria and PERCIST had early predictive value to long-term outcome, but moderate value for pCR. Furthermore, PERCIST might show more potential than the EORTC criteria and conventional PET-based parameters to predict prognosis in breast cancer patients following two cycles of neoadjuvant chemotherapy.Video abstract: see http://links.lww.com/NMC/A162.
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Neoplasias de la Mama/terapia , Terapia Neoadyuvante/métodos , Adulto , Anciano , Neoplasias de la Mama/metabolismo , Diagnóstico Precoz , Femenino , Fluorodesoxiglucosa F18 , Glucólisis , Humanos , Estimación de Kaplan-Meier , Persona de Mediana Edad , Recurrencia Local de Neoplasia , Tomografía Computarizada por Tomografía de Emisión de Positrones , Valor Predictivo de las Pruebas , Pronóstico , Supervivencia sin Progresión , Radiofármacos , Criterios de Evaluación de Respuesta en Tumores SólidosRESUMEN
OBJECTIVE: To critically evaluate the currently available randomized clinical trials regarding the effectiveness of acupuncture in palliative care for cancer patients, hence, to provide sufficient evidences for the widespread use of acupuncture in cancer treatment. METHODS: Two independent reviewers extracted data from all of the randomized clinical trials (RCTs) that assessed the efficacy of acupuncture in palliative care for cancer patients. Seven databases were searched from their respective inception to December 2010. All eligible trials identified were evaluated by two independent reviewers using the Jadad scale, and data from the articles were validated and extracted. RESULTS: In total, 33 RCTs met the inclusion criteria. The effects of acupuncture on different cancer-related aspects were shown, including chemotherapy or radiotherapy-induced side effects (13/33, 39.4%), cancer pain (6/33, 18.2%), post-operative urinary retention (4/33, 12.1%), quality of life (2/33, 6.1%), vasomotor syndrome (2/33, 6.1%), post-operative gastrointestinal dysfunction (2/33, 6.1%), prevention of prolonged postoperative ileus (2/33, 6.1%), joint symptoms (1/33, 3.0%), and immunomodulation (1/33, 3.0%). CONCLUSIONS: The result of our systematic review suggested that the effectiveness of acupuncture in palliative care for cancer patients is promising, especially in reducing chemotherapy or radiotherapyinduced side effects and cancer pain. Acupuncture may be an appropriate adjunctive treatment for palliative care.