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PURPOSE: Immunotherapy is an emerging therapeutic modality for many autoimmune, oncology, and infectious diseases to cure or prevent the underlying causes. Several immunotherapeutic agents are investigated for their beneficial potential in patients with diabetes. However, none have culminated into a successful therapy. The present comprehensive meta-analysis and systematic review covers the last two decades of historical research evaluating the Bacillus Calmette-Guerin (BCG) vaccine as an immunotherapeutic agent in diabetes, along with updated information on similar recent publications. METHOD: A total of 278 articles were retrieved through literature databases, and after applying inclusion and exclusion criteria as per PRISMA guidelines, seven studies were selected for meta-analysis using Cochrane Q statistics. RESULTS: Our meta-analysis revealed marginal benefits, lowering glycosylated/glycated haemoglobin (HbA1C) levels and glutamic-acid-decarboxylase (GAD) autoantibodies in BCG treated people with Type 1 Diabetes (T1D) compared to the matched control individuals. The BCG intervention found to be ineffective in regulating C-peptide (connecting peptide) and clinical remission (CR) i.e. improved glycemic regulation, though beneficial tendency was observed. CONCLUSION: Our systematic review and meta-analysis revealed benefits of BCG vaccine intervention in T1D patients, including improved HbA1C and GAD autoantibody levels. However, the study has several limitations stemming from BCG vaccine-related factors and patient characteristics. Therefore, a large clinical trial with an enhanced study design is needed to validate the immunity-related benefits of the BCG vaccine for glucose metabolism in patients with T1D.
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Alveolar type-II cells (ATII cells) are lung progenitor cells responsible for regeneration of alveolar epithelium during homeostatic turnover and in response to injury. Characterization of ATII cells will have a profound impact on our understanding and treatment of lung disease. The identification of novel ATII cell-surface proteins can be used for sorting and enrichment of these cells for further characterization. Here we combined a high-resolution mass spectrometry-based membrane proteomic approach using lungs of the SILAC mice with an Affymetrix microarray-based transcriptome analysis of ATII cells. We identified 16 proteins that are enriched in the membrane fraction of ATII cells and whose genes are highly expressed in these cells. Interestingly, we confirmed our data for two of these genes, integrin beta 2 and 6 (Itgb2 and Itgb6), by qRT-PCR expression analysis and Western blot analysis of protein extracts. Moreover, flow cytometry and immunohistochemistry in adult lung revealed that ITGB2 and ITGB6 are present in subpopulations of surfactant-associated-protein-C-positive cells, suggesting the existence of different types of ATII cells. Furthermore, analysis of the Itgb2(-/-) mice showed that Itgb2 is required for proper WNT signaling regulation in the lung.
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Antígenos CD18/genética , Células Epiteliales/metabolismo , Cadenas beta de Integrinas/genética , Proteoma/genética , Células Madre/citología , Células Madre/metabolismo , Vía de Señalización Wnt/genética , Animales , Antígenos CD18/metabolismo , Membrana Celular/química , Membrana Celular/metabolismo , Células Epiteliales/citología , Regulación de la Expresión Génica , Cadenas beta de Integrinas/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Anotación de Secuencia Molecular , Unión Proteica , Proteína C/genética , Proteína C/metabolismo , Proteoma/metabolismo , Alveolos Pulmonares/citología , Alveolos Pulmonares/metabolismo , Mucosa Respiratoria/citología , Mucosa Respiratoria/metabolismo , Análisis de Matrices TisularesRESUMEN
Bone is one of the most complex, inaccessible body structures, responsible for calcium storage and haematopoiesis. The second highest cause of death across the world is cancer. Amongst all the types of cancers, bone cancer treatment modalities are limited due to the structural complexity and inaccessibility of bones. The worldwide incidence of bone diseases and bone defects due to cancer, infection, trauma, age-related bone degeneration is increasing. Currently different conventional therapies are available for bone cancer such as chemotherapy, surgery and radiotherapy, but they have several disadvantages associated with them. Nanomedicine is being extensively researched as viable therapeutics to mitigate drug resistance in cancer therapy and promote bone regeneration. Several natural polymers such as chitosan, dextran, alginate, hyaluronic acid, and synthetic polymers like polyglycolic acid, poly(lactic-co-glycolic acid), polycaprolactone are investigated for their application in nanomedicine for bone cancer treatment and bone regeneration. Nanocarriers have shown promising results in preclinical experimental studies. However, they still face a major drawback of inadequate targetability. The paper summarizes the status of research and the progress made so far in modifications and functionalization of natural polymers for improving their site specificity and targeting for effective treatment of bone cancer and enhancing bone regeneration.
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Neoplasias Óseas , Regeneración Ósea , Humanos , Ácido Poliglicólico/química , Ácido Poliglicólico/farmacología , Neoplasias Óseas/tratamiento farmacológicoRESUMEN
Fibrocytes are bone marrow-derived monocytic cells implicated in wound healing. Here, we identify their role in lung cancer progression/ metastasis. Selective manipulation of fibrocytes in mouse lung tumor models documents the central role of fibrocytes in boosting niche features and enhancing metastasis. Importantly, lung cancer patients show increased number of circulating fibrocytes and marked fibrocyte accumulation in the cancer niche. Using double and triple co-culture systems with human lung cancer cells, fibrocytes, macrophages and endothelial cells, we substantiate the central features of cancer-supporting niche: enhanced cancer cell proliferation and migration, macrophage activation, augmented endothelial cell sprouting and fibrocyte maturation. Upregulation of endothelin and its receptors are noted, and dual endothelin receptor blockade suppresses all cancer-supportive phenotypic alterations via acting on fibrocyte interaction with the cancer niche. We thus provide evidence for a crucial role of fibrocytes in lung cancer progression and metastasis, suggesting targets for treatment strategies.
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Células Endoteliales , Neoplasias Pulmonares , Animales , Endotelinas , Fibroblastos/patología , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Ratones , Monocitos/patología , Receptores de EndotelinaRESUMEN
Fibrocytes comprise a recently described cell type of blood-derived, fibroblast-like cells that are recruited from the circulation to sites of wound repair, vascular remodeling, or fibrotic tissue remodeling. We recently showed that the stable prostacyclin analogue treprostinil, a clinically approved drug for pulmonary arterial hypertension (PAH), significantly reduced the recruitment of fibrocytes to sites of vascular remodeling in experimental hypoxic pulmonary hypertension. Here we report on the molecular mechanism underlying the inhibitory action of treprostinil on the adhesion and differentiation of human fibrocytes. Human fibrocytes expressed the prostanoid receptors, prostaglandin I (IP) receptors and prostaglandin E subtype receptors (EP2 and EP4). The generation of intracellular cyclic adenosine monophosphate (cAMP) by treprostinil reduced the expression of the integrins CD49 and CD29 when freshly isolated human peripheral blood mononuclear cells were treated with treprostinil. Cell-matrix adhesion was significantly impaired by treatment with treprostinil. We present evidence for a treprostinil/cAMP-induced downstream suppression of extracellular regulated kinase (ERK) that is transmitted via a protein kinase A-independent pathway through Rap proteins, which sequester Ras. The resulting dephosphorylated state of c-Raf limits the activity of ERK. The cell-matrix adhesion assay with the ERK inhibitor further confirmed that the adhesion of fibrocytes was impaired. Thus our data suggest that treprostinil inhibits the adhesion and differentiation of fibrocytes by limiting the activity of ERK via the cAMP-Rap axis.
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Adhesión Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , AMP Cíclico/metabolismo , Epoprostenol/análogos & derivados , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Leucocitos Mononucleares/efectos de los fármacos , Inhibidores de Proteínas Quinasas/farmacología , Proteínas de Unión al GTP rap1/metabolismo , Proteínas ras/metabolismo , Células Cultivadas , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Epoprostenol/farmacología , Matriz Extracelular/metabolismo , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Hipertensión Pulmonar Primaria Familiar , Humanos , Hipertensión Pulmonar/enzimología , Integrina alfa5/metabolismo , Integrina beta1/metabolismo , Leucocitos Mononucleares/enzimología , Mutación , Fosforilación , Proteínas Proto-Oncogénicas c-raf/metabolismo , Receptores de Prostaglandina/metabolismo , Transducción de Señal/efectos de los fármacos , Factores de Tiempo , Transfección , Proteínas de Unión al GTP rap1/genéticaRESUMEN
AIM: The aim of this study is to evaluate silk-fibroin electrospun nanofibers and blood-derived fibroblast-like cells for cytotoxicity and cell adhesion. BACKGROUND: Silk fibroin (SF) has emerged as a favorable and potential bio-material owing to its unique properties such as biocompatibility, biodegradability, the possibility of functional modifications, mechanical strength, and regenerative capability. Despite current advancements in tissue engineering technologies, delay wound healing and scar formation remain unresolved. Bioequivalent skin graft having human fibroblast and keratinocytes (Apligraft®) has proven to be beneficial, but the cost is a limiting factor. OBJECTIVE: The blood born fibroblast-like cells express several growth factors, extracellular matrix proteins, and these factors are crucial in the various steps of the wound-healing process. SF is an idea polymer by the virtue of its multifaceted characteristics such as mechanical strength, biodegradability, improved cell attachment, biocompatibility, good elasticity, having application in biomedical, tissue engineering, and medicine. The objective of the present study is to evaluate SF as a biomaterial for making nanofibers scaffold and culturing blood-derived fibroblast-like cells on it for the potential application to wound site. MATERIALS AND METHODS: Blood-derived fibroblast-like cells evaluated for cytotoxicity, collagen 1 expression, and cell adhesion on SF electrospun nanofibers. The silk nanofibers were fabricated by the electrospinning method using silk-derived fibroin solution and analyzed for protein composition, viscosity, and further characterized using the Fourier transformed infrared spectroscopy. RESULTS: The SF nanofibers were nontoxic to the blood-derived fibroblast-like cells. It improved cell adhesion with collagen 1 expression. CONCLUSION: The composite scaffold of SF nanofibers with blood-derived fibroblast-like cells would be a potential healing patch for many types of wounds.
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Adhesión Celular/efectos de los fármacos , Fibroblastos/citología , Fibroínas/farmacología , Cicatrización de Heridas , Humanos , Nanofibras , Seda , Ingeniería de TejidosRESUMEN
During the last two decades, the development in drug discovery is slackening due to drug withdrawal from the market or reported to have postmarket safety events. The vital organ toxicities, especially cardiotoxicity, hepatotoxicity, pulmonary toxicity, and neurotoxicity are the major concerns for high drug attrition rates. The pharmaceutical industry is looking for high throughput, high content analysis based novel assays that would be fast, efficient, reproducible, and cost-effective; would address toxicity, the safety of lead molecules, and complement currently used cell-based assays in preclinical testing. The use of zebrafish, a vertebrate screening model, for preclinical testing is increasing owing to the number of advantages and striking similarities with the mammal. The zebrafish embryo development is fast and all vital organs such as the heart, liver, brain, pancreas, and kidneys in zebrafish are functional within 96-120hpf. The maintenance cost of zebrafish is reasonably low as compared to mammalian systems. Due to these features, zebrafish has arisen as a potential experimental screening model in lead identification and validation in the drug efficacy, toxicity, and safety evaluation. Numbers of drugs and chemicals are screened using zebrafish embryos, and results were found to show 100% concordance with mammalian screening data. The application of zebrafish, being a whole-organism screening model, would show a significant reduction in the cost and time required in the drug development process. The present challenge includes complete automation of the zebrafish screening model, i.e., from sorting, imaging of embryos to data analysis to accelerate the therapeutic target identification, and validation process.
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Evaluación Preclínica de Medicamentos , Pez Cebra , Animales , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Dosificación Letal Mediana , Modelos Animales , Pruebas de ToxicidadRESUMEN
BACKGROUND AND PURPOSE: Recruitment and involvement of bone-/blood-derived circulating fibrocytes (CF) in the promotion of fibrotic tissue remodelling processes have been shown. However, their direct contribution to pathological changes is not clear. The present study investigates the causal role of CF in the pathogenesis of pulmonary hypertension (PH). EXPERIMENTAL APPROACH: For selective ablation of CF, we applied the suicidal gene strategy with herpes simplex virus thymidine kinase (HSV-TK) and ganciclovir. The transgenic mice were generated, having HSV-TK-GFP transgene under the collagen 1 promoter. To selectively target CF, HSV-TK-GFP+ bone marrow transplanted into irradiated wild type mice. These chimera mice were subjected to hypoxia for PH induction and ganciclovir for CF ablation. KEY RESULTS: In vivo CF ablation reduced right ventricular hypertrophy and vascular remodelling with reduced total collagen content. We quantified the CF recruited in the perivascular area and arterial wall of small pulmonary arteries. There was significant recruitment of CF in the lung in response to hypoxia. The characterization of CF showed the expression of CD45 and collagen1 (GFP) along with α-smooth muscle actin (αSMA). CONCLUSION AND IMPLICATIONS: Our data demonstrated that CF ablation has a potential impact on right ventricular hypertrophy and vascular remodelling in the setting of experimental pulmonary hypertension induced by hypoxia. The beneficial effects may be related to the direct contribution of fibrocytes or its paracrine effect on other resident cell types. Thus, clinical manipulation of CF may represent a novel therapeutic approach to ameliorate the disease state in pulmonary hypertension.