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The use of tyrosine kinase inhibitors (TKI) has been growing in veterinary oncology and in the past few years several TKI have been tested in dogs. However, different from human medicine, we lack strategies to select patients to be treated with each TKI. Therefore, this study aimed to screen different tumor subtypes regarding TKI target immunoexpression as a predictor strategy to personalize the canine cancer treatment. It included 18 prostatic carcinomas, 36 soft tissue sarcomas, 20 mammary gland tumors, 6 urothelial bladder carcinomas, and 7 tumors from the endocrine system. A total of 87 patients with paraffin blocks were used to perform immunohistochemistry (IHC) of human epidermal growth factor receptor 2 (HER-2), epidermal growth factor receptors 1 (EGFR1), vascular endothelial growth factor receptor 2 (VEGFR-2), platelet derived growth factor receptor beta (PDGFR-ß), c-KIT, and extracellular signal-regulated kinase 1/2 (ERK1/ERK2). The immunohistochemical screening revealed a heterogeneous protein expression among histological types with mesenchymal tumors showing the lowest expression level and carcinomas the highest expression. We have demonstrated by IHC screening that HER2, EGFR1, VEGFR-2, PDGFR-ß and ERK1/ERK2 are commonly overexpressed in dogs with different carcinomas, and KIT expression is considered relatively low in the analyzed samples.
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Enfermedades de los Perros , Inmunohistoquímica , Perros , Animales , Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/patología , Masculino , Femenino , Neoplasias/metabolismo , Neoplasias/tratamiento farmacológico , Neoplasias/veterinaria , Neoplasias/patología , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Proteínas Tirosina Quinasas Receptoras/metabolismo , Inhibidores de Proteínas Quinasas/uso terapéutico , Inhibidores de Proteínas Quinasas/farmacología , Biomarcadores de Tumor/metabolismo , Receptor ErbB-2/metabolismo , Proteínas Proto-Oncogénicas c-kit/metabolismo , Receptores ErbB/metabolismo , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo , HumanosRESUMEN
The present study describes the morphological and immunohistochemical characteristics of a case of diffuse seminoma in a 16-year-old male mixed-breed horse. According to the owner, the animal's left testicle had been gradually increasing in size over a period of 2 months. On palpation, the testicle had a firm consistency, with no sensitivity to digital pressure, was adhered to the scrotum and measuring 16 cm × 8 cm. In the ultrasound examination, it presented a heterogeneous texture and areas of hypoechogenic echogenicity without visualization of the mediastinum. Therefore, the bilateral orchiectomy was performed. After the surgical procedure, it was found that the affected testicle presented a firm mass measuring 9 cm × 7 cm × 3.5 cm. Histologically, a multilobulated, non-encapsulated and invasive tumour mass was found, which replaced the seminiferous tubules, consisting of polygonal cells arranged in a mantle that varied from cohesive to loosely cohesive, supported by a scarce fibrous stroma. In the immunohistochemical examination, the neoplastic cells showed positive immunolabelling for OCT4 and C-KIT. In this report, the physical examination combined with the ultrasonographic examination were fundamental to the therapeutic management of the case, and the final diagnosis was made after histopathological and immunohistochemical tests.
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Enfermedades de los Caballos , Orquiectomía , Seminoma , Neoplasias Testiculares , Masculino , Animales , Seminoma/veterinaria , Seminoma/patología , Seminoma/cirugía , Caballos , Neoplasias Testiculares/veterinaria , Neoplasias Testiculares/patología , Neoplasias Testiculares/cirugía , Orquiectomía/veterinaria , Enfermedades de los Caballos/patología , Enfermedades de los Caballos/cirugía , Enfermedades de los Caballos/diagnóstico por imagen , Ultrasonografía/veterinaria , Inmunohistoquímica/veterinaria , Proteínas Proto-Oncogénicas c-kit/análisis , Proteínas Proto-Oncogénicas c-kit/metabolismo , Factor 3 de Transcripción de Unión a Octámeros/análisis , Factor 3 de Transcripción de Unión a Octámeros/metabolismoAsunto(s)
Mastocitosis Sistémica , Mastocitosis , Humanos , Mastocitos/metabolismo , Mastocitos/patología , Brasil , Centros de Atención Terciaria , Mastocitosis/diagnóstico , Mastocitosis/metabolismo , Mastocitosis/patología , Biopsia , Algoritmos , Mastocitosis Sistémica/diagnóstico , Proteínas Proto-Oncogénicas c-kitRESUMEN
This study aims to define the best method (slow freezing or vitrification) and fragment size (1, 5, or 9 mm³) for prepubertal goat testis cryopreservation, as well as to evaluate testicular morphological integrity after cryopreservation and in vitro culture (IVC). Initially (experiment I), 1, 5, or 9 mm³ testis fragments were cryopreserved by slow freezing using a Mr. Frosty container with 20% Dimethylsulfoxide (DMSO) or vitrified using the Ovarian Tissue Cryosystem (OTC) device, (Equilibration solution - ES: 10% DMSO and 10% ethylene glycol - EG; Vitrification solution - VS: 20% DMSO and 20% EG) and then subjected to morphological analysis, type I and III collagen quantification and gene expression (Oct4, C-kit, Bax, and Bcl-2). Subsequently, (experiment II), fresh or cryopreserved by slow freezing testis fragments were cultured in vitro and submitted to morphological analysis by scanning electron microscopy. The data from the experiment I revealed fewer morphological alterations in 1 and 5 mm³ fragments after vitrification and slow freezing, respectively. The percentage of type I collagen fibers in 5 and 9 mm³ frozen was higher than in fresh or vitrified fragments. For type III collagen, fresh or frozen fragments of 1 and 5 mm3 showed a higher percentage than fragments of 9 mm3. Gene expression for Oct4 and C-kit after slow freezing or vitrification in the 5 mm3 fragments was lower than that observed in the fresh fragments. The Bax:Bcl-2 ratio in the 1 and 9 mm³ fragments was lower than in the 5 mm³ fragments for fresh fragments or after freezing. In experiment II, fragments cultured in vitro, previously frozen or not, showed more morphological alterations than fresh or frozen fragments. We concluded that slow freezing of 5 mm³ fragments was the best protocol for cryopreserving prepubertal goat testis and although the results of IVC are encouraging, it still needs improvement to restore testicular function after cryopreservation.
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Dimetilsulfóxido , Cabras , Animales , Masculino , Proteína X Asociada a bcl-2 , Criopreservación/veterinaria , Proteínas Proto-Oncogénicas c-bcl-2 , Proteínas Proto-Oncogénicas c-kitRESUMEN
Embora o melanoma seja representado por apenas cerca de 5% dos casos de tumores malignos de pele, é responsável pela ampla maioria de óbitos relacionados a tumores cutâneos. Seu desenvolvimento está principalmente associado à presença de mutações em oncogenes específicos, como BRAF, c-KIT e PDGFRA, os quais estão relacionados ao metabolismo celular. OBJETIVOS: Avaliar retrospectivamente a presença de mutações nos genes BRAF (éxons 15), C-KIT (éxons 9,11,13 e 17) e PDGFRA (éxons 12 ,14 e 18) e correlacionar os resultados com as características clínicas e epidemiológicas e desfecho dos pacientes. METODOLOGIA: Foi realizado um estudo retrospectivo de 94 pacientes com melanoma metastático atendidos no Hospital Amaral Carvalho entre os anos de 2015 a 2022, de acordo com as seguintes variáveis: idade ao diagnóstico, gênero, cor, local do tumor primário, local de metástase, subtipo histológico de melanoma, dosagem de LDH, positividade para marcadores de imuno-histoquímica, espessura, presença de ulceração, metástase no linfonodo ao diagnóstico do tumor primário, espessura Breslow, classificação Clark, índice mitótico e desfecho (óbito, sobrevida). RESULTADOS: A maioria dos pacientes eram do sexo masculino (52,1%), de cor branca (97,9%) e com idade acima de 60 anos (42,5%). O subtipo histológico mais encontrado foi o extensivo superficial (42,5%), sendo os membros inferiores o local mais acometido (35,1%). O nível IV de Clark (39,4%) e índice de Breslow acima de 4mm (33,0%) foram os mais prevalentes. A maior parte dos tumores não apresentou ulceração (53,2%) e mais da metade dos pacientes apresentou mais de uma mitose por mm² (47,9%). A dosagem de desidrogenase láctica (LDH) se manteve dentro da normalidade na maioria dos pacientes no momento do diagnóstico (43,6%) e no surgimento de metástases (30,9%). Todos os pacientes apresentaram ao menos uma metástase, sendo os linfonodos regionais o local de maior incidência. A maioria dos pacientes faleceu em decorrência da doença (66,0%). A análise de BRAF apresentou significância estatística para as variáveis idade ao diagnóstico (p= 0,0085), tipo histológico (p= 0,048) e nível de Clark (p= 0,0290). A mediana de sobrevivência para pacientes com mutações em BRAF foi de 62 meses e 50 meses para aqueles que não apresentaram mutações. Não foram encontradas significâncias estatísticas entre c-KIT e as variáveis analisadas. A mediana de sobrevivência para pacientes com mutações em c-KIT foi de 38 meses e 70 meses para aqueles que não apresentaram mutações. CONCLUSÕES: A presença de mutação em BRAF esteve associada à menor idade e menores índices de Clark, e relacionada à ligeira maior sobrevida. O contrário foi observado para mutações em c-KIT, apesar da ausência de significância estatística: uma ligeira menor sobrevida foi observada em pacientes mutados. O LDH não esteve bem correlacionado à presença do melanoma metastático ao se apresentar normal na maioria dos casos. Grande parte dos pacientes tiveram o tumor primário localizado nos membros inferiores, diferentemente do relatado por outros estudos, e apesar de não significante, os tumores de tronco foram em sua maioria mutados para BRAF, e o oposto observado em membros inferiores e superiores. Assim, o papel prognóstico da mutação em BRAF na progressão da doença é ainda controverso. Tais dados podem fornecer mais informações para a adequação de protocolos de prevenção e tratamento de pacientes acometidos por melanoma.
INTRODUCTION: Although melanoma is represented by only about 5% of cases of malignant skin tumors, it is responsible for the vast majority of deaths related to skin tumors. Its development is mainly associated with the presence of mutations in specific oncogenes, such as BRAF, c-KIT and PDGFRA, which are related to cell metabolism. OBJECTIVES: Retrospectively evaluate the presence of mutations in BRAF (exons 15), C-KIT (exons 9,11,13 and 17) and PDGFRA (exons 12, 14 and 18) genes and correlate the results with the clinical and epidemiological characteristics and patient outcomes. METHODS: A retrospective study of 94 patients with metastatic melanoma treated at Hospital Amaral Carvalho between the years 2015 to 2022 was carried out, according to the following variables: age at diagnosis, gender, color, site of primary tumor, site of metastasis, histological subtype of melanoma, LDH dosage, positivity for immunohistochemical markers, thickness, presence of ulceration, lymph node metastasis at diagnosis of the primary tumor, Breslow thickness, Clark classification, mitotic index and outcome (death, survival). RESULTS: Most patients were male (52.1%), white (97.9%) and aged over 60 years (42.5%). The most common histological subtype was superficial extensive (42.5%), with the lower limbs being the most affected site (35.1%). Clark's level IV (39.4%) and Breslow thickness above 4mm (33.0%) were the most prevalent. Most tumors did not show ulceration (53.2%) and more than half of the patients had more than one mitosis per mm² (47.9%). The lactate dehydrogenase (LDH) dosage remained within the normal range in most patients at the time of diagnosis (43.6%) and at the onset of metastases (30.9%). All patients had at least one metastasis, with regional lymph nodes being the site with the highest incidence. Most patients died as a result of the disease (66.0%). BRAF analysis showed statistical significance for the variables age at diagnosis (p= 0.0085), histological type (p= 0.048) and Clark's level (p= 0.0290). Median survival for patients with BRAF mutations was 62 months and 50 months for those without mutations. No statistical significance was found between c-KIT and the analyzed variables. The median survival for patients with c-KIT mutations was 38 months and 70 months for those without mutations. CONCLUSIONS: The presence of a BRAF mutation was associated with younger age and lower Clark scores, and related to slightly longer survival. The opposite was observed for mutations in c-KIT, despite the absence of statistical significance: a slightly lower survival was observed in mutated patients. LDH was not well correlated with the presence of metastatic melanoma as it was normal in most cases. Most of the patients had the primary tumor located in the lower limbs, unlike what has been reported by other studies, and although not significant, the trunk tumors were mostly mutated to BRAF, and the opposite was observed in the lower and upper limbs. Thus, the prognostic role of the BRAF mutation in disease progression is still controversial. Such data may provide more information for the adequacy of prevention and treatment protocols for patients affected by melanoma.
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Humanos , Masculino , Femenino , Proteínas Proto-Oncogénicas c-kit , Proteínas Proto-Oncogénicas B-raf , Melanoma/metabolismo , Mutación , Neoplasias Cutáneas , Instituciones Oncológicas/estadística & datos numéricos , Estudios Retrospectivos , Epidemiología Clínica , Melanoma/genéticaRESUMEN
Triple-negative breast cancer (TNBC) is associated with a poor prognosis and the absence of targeted therapy. c-Kit, a receptor tyrosine kinase (RTK), is considered a molecular target for anticancer drugs. Tyrosine kinase inhibitors (TKIs) recognizing c-Kit are used for the treatment of c-Kit-expressing tumors. However, the expression, function, and therapeutic potential of c-Kit have been little explored in TNBC. Here, we studied the expression and effects of c-Kit in TNBC through in vitro and in silico analysis, and evaluated the response to TKIs targeting c-Kit. Analysis of TNBC cells showed the expression of functional c-Kit at the cell membrane. The stimulation of c-Kit with its ligand induced the activation of STAT3, Akt, and ERK1/2, increasing cell migration, but had no effect on cell proliferation or response to Doxorubicin. Analysis of public datasets showed that the expression of c-Kit in tumors was not associated with patient survival. Finally, TNBC cells were susceptible to TKIs, in particular the effect of Nilotinib was stronger than Doxorubicin in all cell lines. In conclusion, TNBC cells express functional c-Kit, which is a targetable molecule, and show a strong response to Nilotinib that may be considered a candidate drug for the treatment of TNBC.
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Neoplasias de la Mama Triple Negativas , Línea Celular Tumoral , Proliferación Celular , Doxorrubicina/farmacología , Humanos , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Proteínas Proto-Oncogénicas c-kit/genética , Proteínas Tirosina Quinasas Receptoras , Neoplasias de la Mama Triple Negativas/patologíaRESUMEN
Mast cell tumors (MCTs) are an uncommon primary neoplasm of the nasal cavity in dogs for which there is a paucity of existing literature regarding their clinical behavior and molecular features. The objectives of this retrospective study were to examine the clinical findings, histopathologic and immunohistochemical features, and c-KIT mutation status of primary intranasal MCTs in dogs and identify potential prognostic factors. Canine biopsies submitted to a diagnostic laboratory in Colorado between 2010 and 2019 with intranasal neoplasms diagnosed as MCTs and no history of cutaneous or oral MCT were considered. Immunohistochemistry for CD117 and Ki67 and polymerase chain reaction (PCR) for internal tandem duplications at exons 8 and 11 of the c-KIT gene were performed. Twenty out of 1849 (1%) primary intranasal neoplasms were MCTs. Metastases were reported in 11/20 cases (55%), with the mandibular lymph node representing the most common site. One case had distant metastases to abdominal viscera. Of the cases with available outcome data, 6/14 (43%) died or were euthanized from MCT-related disease within 1 year of the onset of clinical signs. Only one case had a c-KIT mutation at exon 11. In our study, intranasal MCTs were prone to metastasize and had a generally poor prognosis, resembling the behavior of MCTs arising in other mucosal locations. While dogs with metastatic disease and survival times of <1 year tended to have atypical KIT localization, moderate to high Ki67 indices, and mitotic counts ≥8, definitive prognosticators could not be identified due to the limited number of cases with favorable clinical outcomes.
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Enfermedades de los Perros , Neoplasias Cutáneas , Animales , Enfermedades de los Perros/patología , Perros , Antígeno Ki-67 , Mastocitos/patología , Proteínas Proto-Oncogénicas c-kit/genética , Estudios Retrospectivos , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/veterinariaRESUMEN
Systemic mastocytosis (SM) is a myeloid neoplasm characterized by abnormal growth and accumulation of neoplastic mast cells in at least one extracutaneous site. Clinical presentation and course are variable, most patients are developing an indolent disease and some, an aggressive/leukemic form. Because of its rarity, most physicians are unfamiliar with this disease and do not readily diagnose it. In the present retrospective study, the authors describe 12 patients diagnosed with mast cell neoplasm. Cases were selected from three institutions from Campinas and São Paulo City, Brazil. Morphological features and diagnostic pitfalls are emphasized. Patients' age ranged from 15 to 81 years (mean 51.6 years). Male and female were affected similarly (1:1). Ten patients were classified as aggressive SM, one patient as SM with an associated acute promyelocytic leukemia with t(15;17), and one patient with mast cell sarcoma. The most common clinical findings included anemia (9 patients), thrombocytopenia (3 patients), and skin lesions (3 patients). Bone marrow was involved in 11 patients at diagnosis, followed by skin (5 patients). Five morphological patterns were present: mast cell aggregates (5), plasmacytoid (4), monocytoid (2), spindle cell (2), and epithelioid/pleomorphic (1); two patients showed two histological patterns. In all cases, neoplastic cells were positive for CD117/C-KIT. C-KIT D816V mutation was present in four patients, C-KIT K509I in two, and del(7q22) in one; in five cases no mutational status was available. Despite limited resources, basically morphology and a restricted immunohistochemical panel, it is possible to diagnose mast cell neoplasm. Of note, the pathologist should recognize the different morphological variants of the disease and include adequate markers when requesting immunohistochemical studies.
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Mastocitosis Sistémica , Trastornos Mieloproliferativos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Brasil , Femenino , Humanos , Masculino , Mastocitos/patología , Mastocitosis Sistémica/diagnóstico , Mastocitosis Sistémica/genética , Mastocitosis Sistémica/patología , Persona de Mediana Edad , Trastornos Mieloproliferativos/diagnóstico , Trastornos Mieloproliferativos/genética , Proteínas Proto-Oncogénicas c-kit/análisis , Proteínas Proto-Oncogénicas c-kit/genética , Estudios Retrospectivos , Adulto JovenRESUMEN
ABSTRACT: Melanomas that arise in sun-protected sites, including acral and oral mucosal melanomas, are likely under the control of unique, specific mechanisms that lead to mutagenesis through various pathways. In this study, we examined somatic mutations in tumors by targeted sequencing using a custom Ion Ampliseq Panel, comprising hotspots of 14 genes that are frequently mutated in solid tumors. Tumor DNA was extracted from 9 formalin fixation, paraffin-embedded sun-protected melanomas (4 primary oral mucosal melanomas and 5 acral lentiginous melanomas), and we identified mutations in the NRAS , PIK3CA , EGFR , HRAS , ERBB2 , and ROS1 genes. This study reveals new actionable mutations that are potential targets in the treatment of photo-protected melanomas. Additional studies on more of these melanoma subtypes could confirm our findings and identify new mutations.
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Melanoma , Neoplasias Cutáneas , Fosfatidilinositol 3-Quinasa Clase I/genética , Formaldehído , Humanos , Melanoma/patología , Mutación , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas B-raf/genética , Proteínas Proto-Oncogénicas c-kit/genética , Neoplasias Cutáneas/patologíaRESUMEN
Gastrointestinal stromal tumors (GISTs) are unusual cancers, which are developed in specialized cells in the gastrointestinal tract wall. Various strategies involving single-agents, combinations, and rapid complementary inhibitor cycling are now being used to control such tumors. Based on promising early clinical trial experience, certain novel KIT and PDGFRA tyrosine kinase inhibitors have shown advanced clinical development. Resistance to tyrosine kinase inhibitors has brought immense difficulties, with patients now requiring additional therapeutic options. This review describes and discusses the last five years (2016-2020) in developing novel c-KIT kinase inhibitors using virtual screening and docking approaches. Computational techniques can be used to complement experimental studies to identify new candidate molecules for therapeutic use. Molecular modeling strategies allow the analysis of the required characteristics that compounds must have to effectively bind c-KIT. Through such analyses, it is possible to both discover and design novel inhibitors against cancer-related proteins that play a critical role in tumor development (including mutant strains). Docking showed potential in the detection of the key residues responsible for ligand recognition and is very helpful to understand the interactions in the active site that can be used to develop new compounds/classes of anticancer drugs and help millions of cancer patients.
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Antineoplásicos , Tumores del Estroma Gastrointestinal , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Tumores del Estroma Gastrointestinal/tratamiento farmacológico , Humanos , Simulación del Acoplamiento Molecular , Mutación , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Proteínas Proto-Oncogénicas c-kit/genética , Proteínas Proto-Oncogénicas c-kit/uso terapéuticoRESUMEN
Innate lymphoid cells (ILCs) are the most recently described group of lymphoid subpopulations. These tissue-resident cells display a heterogeneity resembling that observed on different groups of T cells, hence their categorization as cytotoxic NK cells and helper ILCs type 1, 2 and 3. Each one of these groups is highly diverse and expresses different markers in a context-dependent manner. Type 2 innate lymphoid cells (ILC2s) are activated in response to helminth parasites and regulate the immune response. They are involved in the etiology of diseases associated with allergic responses as well as in the maintenance of tissue homeostasis. Markers associated with their identification differ depending on the tissue and model used, making the study and understanding of these cells a cumbersome task. This review compiles evidence for the heterogeneity of ILC2s as well as discussion and analyses of molecular markers associated with their identity, function, tissue-dependent expression, and how these markers contribute to the interaction of ILC2s with specific microenvironments to maintain homeostasis or respond to pathogenic challenges.
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Antígenos de Diferenciación/análisis , Subgrupos Linfocitarios/inmunología , Tejido Adiposo Blanco/inmunología , Tejido Adiposo Blanco/patología , Animales , Citocinas/metabolismo , Helmintiasis/inmunología , Antígenos de Histocompatibilidad Clase II/inmunología , Homeostasis , Humanos , Inmunofenotipificación , Inflamación , Intestinos/inmunología , Pulmón/inmunología , Subgrupos Linfocitarios/química , Ratones , Nutrientes , Especificidad de Órganos , Proteínas Proto-Oncogénicas c-kit/inmunología , Receptores de Superficie Celular/inmunología , Piel/inmunología , Factor de Células Madre/inmunologíaRESUMEN
Supplements containing pharmacological concentrations of biotin are commercially available. The mechanisms by which biotin at pharmacological concentrations exerts its action have been the subject of multiple investigations, particularly for biotin's medicinal potential and wide use for cosmetic purposes. Several studies have reported that biotin supplementation increases cell proliferation; however, the mechanisms involved in this effect have not yet been characterized. In a previous study, we found that a biotin-supplemented diet increased spermatogonia proliferation. The present study was focused on investigating the molecular mechanisms involved in biotin-induced testis cell proliferation. Male BALB/cAnNHsd mice were fed a control or a biotin-supplemented diet (1.76 or 97.7 mg biotin/kg diet) for eight weeks. Compared with the control group, the biotin-supplemented mice presented augmented protein abundance of the c-kit-receptor and pERK1/2Tyr204 and pAKTSer473, the active forms of ERK/AKT proliferation signaling pathways. No changes were observed in the testis expression of the stem cell factor and in the serum levels of the follicle-stimulating hormone. Analysis of mRNA abundance found an increase in cyclins Ccnd3, Ccne1, Ccna2; Kinases Cdk4, Cdk2; and E2F; and Sp1 & Sp3 transcription factors. Decreased expression of cyclin-dependent kinase inhibitor 1a (p21) was observed but not of Cdkn2a inhibitor (p16). The results of the present study identifies, for the first time, the mechanisms associated with biotin supplementation-induced cell proliferation, which raises concerns about the effects of biotin on male reproductive health because of its capacity to cause hyperplasia, especially because this vitamin is available in large amounts without regulation.
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Biotina/toxicidad , Proliferación Celular/efectos de los fármacos , Suplementos Dietéticos/toxicidad , Hormona Folículo Estimulante/sangre , Espermatogonias/efectos de los fármacos , Factor de Células Madre/metabolismo , Testículo/efectos de los fármacos , Animales , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Masculino , Ratones Endogámicos BALB C , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-kit/genética , Proteínas Proto-Oncogénicas c-kit/metabolismo , Transducción de Señal , Factor de Transcripción Sp1/genética , Factor de Transcripción Sp1/metabolismo , Factor de Transcripción Sp3/genética , Factor de Transcripción Sp3/metabolismo , Espermatogonias/metabolismo , Espermatogonias/patología , Testículo/metabolismo , Testículo/patologíaRESUMEN
BACKGROUND: Human endometrium harbors stem/progenitor cells (SPCs) that may contribute to the establishment of endometriosis when seeded outside the uterus. Oct-4, C-kit and Musashi-1 are some of the many proteins used to characterize SPCs, but their association with endometriosis is uncertain. OBJECTIVE AND DESIGN: In this study, specimens of normal endometrium (n = 12), eutopic endometrium from women with endometriosis (n = 9), superficial peritoneal endometriosis (SUP, n = 12) and deep endometriosis (DE, n = 13) lesions were evaluated for localization and intensity of immunostaining for Oct-4, C-kit and Musashi-1. RESULTS: The three markers were abundantly expressed in normal endometrium, eutopic endometrium from endometriosis patients, SUP and DE specimens. Oct-4 and C-kit expression did not vary across groups as regards intensity or frequency. C-kit staining signal was seldom detected in vascular endothelium of normal or eutopic endometrium from endometriosis patients; however, it was positive in 67% of the SUP lesions and in 25% of the DE lesions (p = 0.042). Musashi-1 was expressed in some endometriotic glands as cell clusters, but its signal was similar between the four types of tissue (p = 0.971) CONCLUSION: The wide distribution of Oct-4, C-kit and Musashi-1 in endometria of patients with and without endometriosis and in SUP and DE endometriotic lesions suggests that these markers are not suitable for the in situ characterization of endometrial SPCs and should not be taken as surrogates for the study of SPCs in the pathogenesis of endometriosis.
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Endometriosis/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Factor 3 de Transcripción de Unión a Octámeros/metabolismo , Proteínas Proto-Oncogénicas c-kit/metabolismo , Proteínas de Unión al ARN/metabolismo , Células Madre/metabolismo , Adulto , Biomarcadores/metabolismo , Biopsia , Endometriosis/patología , Femenino , Humanos , Inmunohistoquímica , Persona de Mediana EdadRESUMEN
Degeneration of motor neurons, glial cell reactivity, and vascular alterations in the CNS are important neuropathological features of amyotrophic lateral sclerosis (ALS). Immune cells trafficking from the blood also infiltrate the affected CNS parenchyma and contribute to neuroinflammation. Mast cells (MCs) are hematopoietic-derived immune cells whose precursors differentiate upon migration into tissues. Upon activation, MCs undergo degranulation with the ability to increase vascular permeability, orchestrate neuroinflammation and modulate the neuroimmune response. However, the prevalence, pathological significance, and pharmacology of MCs in the CNS of ALS patients remain largely unknown. In autopsy ALS spinal cords, we identified for the first time that MCs express c-Kit together with chymase, tryptase, and Cox-2 and display granular or degranulating morphology, as compared with scarce MCs in control cords. In ALS, MCs were mainly found in the niche between spinal motor neuron somas and nearby microvascular elements, and they displayed remarkable pathological abnormalities. Similarly, MCs accumulated in the motor neuron-vascular niche of ALS murine models, in the vicinity of astrocytes and motor neurons expressing the c-Kit ligand stem cell factor (SCF), suggesting an SCF/c-Kit-dependent mechanism of MC differentiation from precursors. Mechanistically, we provide evidence that fully differentiated MCs in cell cultures can be generated from the murine ALS spinal cord tissue, further supporting the presence of c-Kit+ MC precursors. Moreover, intravenous administration of bone marrow-derived c-Kit+ MC precursors infiltrated the spinal cord in ALS mice but not in controls, consistent with aberrant trafficking through a defective microvasculature. Pharmacological inhibition of c-Kit with masitinib in ALS mice reduced the MC number and the influx of MC precursors from the periphery. Our results suggest a previously unknown pathogenic mechanism triggered by MCs in the ALS motor neuron-vascular niche that might be targeted pharmacologically.
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Esclerosis Amiotrófica Lateral/inmunología , Mastocitos/inmunología , Microvasos/patología , Neuronas Motoras/patología , Enfermedades Neuroinflamatorias/inmunología , Proteínas Proto-Oncogénicas c-kit/metabolismo , Médula Espinal/inmunología , Anciano , Anciano de 80 o más Años , Esclerosis Amiotrófica Lateral/metabolismo , Esclerosis Amiotrófica Lateral/patología , Animales , Astrocitos/metabolismo , Astrocitos/patología , Benzamidas/farmacología , Estudios de Casos y Controles , Quimasas/metabolismo , Ciclooxigenasa 2/metabolismo , Femenino , Humanos , Masculino , Mastocitos/efectos de los fármacos , Mastocitos/metabolismo , Ratones , Microvasos/metabolismo , Persona de Mediana Edad , Neuronas Motoras/metabolismo , Enfermedades Neuroinflamatorias/metabolismo , Enfermedades Neuroinflamatorias/patología , Piperidinas/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-kit/antagonistas & inhibidores , Piridinas/farmacología , Médula Espinal/efectos de los fármacos , Médula Espinal/metabolismo , Médula Espinal/patología , Factor de Células Madre/metabolismo , Tiazoles/farmacología , Triptasas/metabolismoRESUMEN
Histological grading systems remain cornerstones in the prognosis of canine cutaneous mast cell tumours (MCTs), but the distinct biological behaviour of each tumour often necessitates the use of complementary markers. Although a plethora of immunohistochemical markers have been proposed as prognostic factors, few are presently applied in routine diagnosis. This systematic review and meta-analysis was designed to establish which immunohistochemical markers have verifiable prognostic value for cutaneous MCTs in dogs. A Boolean search of five databases identified 200 articles for screening, of which 73 were selected for full-text assessment and 24 ultimately included in the systematic review. Odds Ratio (OR) was adopted as the summary measure for subsequent meta-analysis but only 15 articles, relating to the immunomarkers Ki-67 (9), KIT (5), and BAX (2), provided either a value for OR or sufficient data to calculate this statistic. Meta-analysis verified that canine cutaneous MCTs with elevated expression of Ki-67 or BAX, as well aberrant immuno-expression of KIT, showed an increased odds of death, with respective OR values of 11.2 (95% CI 6.3-20.0; p < .01), 9.9 (95% CI 1.3-73.6; p = .03), and 4.1 (95% CI 1.1-15.3; p = .03). Despite KIT, Ki67, and BAX arise as suitable prognostic factor for canine MCTs, this study highlighted the lack of important clinical and statistical data in many published articles, rendering it impossible to complete the meta-analysis of several potentially valuable immunohistochemical markers.
Asunto(s)
Enfermedades de los Perros , Mastocitoma Cutáneo , Mastocitosis Cutánea , Neoplasias Cutáneas , Animales , Enfermedades de los Perros/diagnóstico , Perros , Inmunohistoquímica , Antígeno Ki-67 , Mastocitos , Mastocitoma Cutáneo/diagnóstico , Mastocitoma Cutáneo/veterinaria , Mastocitosis Cutánea/diagnóstico , Mastocitosis Cutánea/veterinaria , Pronóstico , Proteínas Proto-Oncogénicas c-kit , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/veterinaria , Proteína X Asociada a bcl-2RESUMEN
Protein malnutrition causes anemia and leukopenia as it reduces hematopoietic precursors and impairs the production of mediators that regulate hematopoiesis. Hematopoiesis occurs in distinct bone marrow niches that modulate the processes of differentiation, proliferation and self-renewal of hematopoietic stem cells (HSCs). Mesenchymal stem cells (MSCs) contribute to the biochemical composition of bone marrow niches by the secretion of several growth factors and cytokines, and they play an important role in the regulation of HSCs and hematopoietic progenitors. In this study, we investigated the effect of protein malnutrition on the hematopoietic regulatory function of MSCs. C57BL/6NTaq mice were divided into control and protein malnutrition groups, which received, respectively, a normal protein diet (12% casein) and a low protein diet (2% casein). The results showed that protein malnutrition altered the synthesis of SCF, TFG-ß, Angpt-1, CXCL-12, and G-CSF by MSCs. Additionally, MSCs from the protein malnutrition group were not able to maintain the lymphoid, granulocytic and megakaryocytic-erythroid differentiation capacity compared to the MSCs of the control group. In this way, the comprehension of the role of MSCs on the regulation of the hematopoietic cells, in protein malnutrition states, is for the first time showed. Therefore, we infer that hematopoietic alterations caused by protein malnutrition are due to multifactorial alterations and, at least in part, the MSCs' contribution to hematological impairment.
Asunto(s)
Células de la Médula Ósea/efectos de los fármacos , Proteínas en la Dieta/administración & dosificación , Hematopoyesis/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Deficiencia de Proteína/metabolismo , Animales , Células de la Médula Ósea/fisiología , Técnicas de Cocultivo , Medios de Cultivo Condicionados , Hematopoyesis/fisiología , Leucocitos Mononucleares/fisiología , Ratones , Proteínas Proto-Oncogénicas c-kit/metabolismo , ARN/efectos de los fármacos , ARN/genética , ARN/metabolismoRESUMEN
Articular chondral lesions, caused either by trauma or chronic cartilage diseases such as osteoarthritis, present very low ability to self-regenerate. Thus, their current management is basically symptomatic, progressing very often to invasive procedures or even arthroplasties. The use of amniotic fluid stem cells (AFSCs), due to their multipotentiality and plasticity, associated with scaffolds, is a promising alternative for the reconstruction of articular cartilage. Therefore, this study aimed to investigate the chondrogenic potential of AFSCs in a micromass system (high-density cell culture) under insulin-like growth factor 1 (IGF-1) stimuli, as well as to look at their potential to differentiate directly when cultured in a porous chitosan-xanthan (CX) scaffold. The experiments were performed with a CD117 positive cell population, with expression of markers (CD117, SSEA-4, Oct-4 and NANOG), selected from AFSCs, after immunomagnetic separation. The cells were cultured in both a micromass system and directly in the scaffold, in the presence of IGF-1. Differentiation to chondrocytes was confirmed by histology and by using immunohistochemistry. The construct cell-scaffold was also analyzed by scanning electron microscopy (SEM). The results demonstrated the chondrogenic potential of AFSCs cultivated directly in CX scaffolds and also in the micromass system. Such findings support and stimulate future studies using these constructs in osteoarthritic animal models.
Asunto(s)
Células Madre Adultas/citología , Cartílago Articular/efectos de los fármacos , Condrogénesis/genética , Osteoartritis/genética , Andamios del Tejido/química , Células Madre Adultas/trasplante , Líquido Amniótico/citología , Cartílago Articular/crecimiento & desarrollo , Cartílago Articular/ultraestructura , Técnicas de Cultivo de Célula , Diferenciación Celular/efectos de los fármacos , Quitosano/farmacología , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Humanos , Factor I del Crecimiento Similar a la Insulina/farmacología , Microscopía Electrónica de Rastreo , Osteoartritis/patología , Osteoartritis/terapia , Polisacáridos Bacterianos/farmacología , Proteínas Proto-Oncogénicas c-kit/genética , Ingeniería de Tejidos/métodosAsunto(s)
Neoplasias Colorrectales/tratamiento farmacológico , Metástasis de la Neoplasia/tratamiento farmacológico , Compuestos de Fenilurea/uso terapéutico , Piridinas/uso terapéutico , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Supervivencia sin Enfermedad , Humanos , Masculino , Persona de Mediana Edad , Mutación , Metástasis de la Neoplasia/patología , Proteínas Proto-Oncogénicas c-kit/metabolismoRESUMEN
Systemic mastocytosis (SM) pathology is extremely rare in canine practice, with insufficient reported data. The knowledge of the clinical behavior of this pathology is scarce. In human medicine, SM has been widely investigated, being defined as a rare hematopoietic disorder by the World Health Organization (2016), within the type of myeloproliferative neoplasms. Herein, we describe a systemic mastocytosis case in a Portuguese Serra-da-Estrela dog, where a cutaneous grade III/high-grade MCT was also diagnosed. The clinical decline of the animal and owner's insistence throughout anamnesis that the dog was markedly different after the cytologic exam performed in another clinic, along with both severe eosinophilia and hepatomegaly, led to the clinical suspicion of SM. The animal passed away 7 days later. Post-morteminvestigation confirmed SM pathology, and a deletion of 15 base pairs change on c-Kit gene exon 11 was identified. Contemplating the low number of cases described in the literature, this publication aims to disclose clinical and laboratory features of rare and poorly described canine SM, taking into consideration human outcomes described in the literature.(AU)
A patologia da mastocitose sistêmica (SM) é extremamente rara na prática clínica canina, com escassos casos descritos na literatura científica. O conhecimento do comportamento clínico desta patologia é mínimo. Na medicina humana, a SM tem sido amplamente investigada, sendo definida como uma doença hematopoiética rara pela Organização Mundial da Saúde (2016), dentro do tipo de neoplasias mieloproliferativas. Descrevemos aqui um caso de mastocitose sistêmica num cão Serra-da-Estrela português, diagnosticado também com um mastocitoma cutâneo grau III / alto grau. O declínio clínico do animal e a insistência do proprietário durante a anamnese de que o cão estava marcadamente diferente após o exame citológico realizado em outra clínica, juntamente com eosinofilia e hepatomegalia graves, levantaram a suspeita clínica de SM. O animal faleceu 7 dias depois. A investigação post-mortem confirmou a patologia SM, e o estudo molecular revelou uma deleção de 15 pares de bases no exon 11 do gene c-Kit. Contemplando o baixo número de casos descritos na literatura, o objetivo desta publicação é divulgar características clínicas e laboratoriais de SM canina, levando em consideração informações clínicas descritas em humanos.(AU)