RESUMO
Vagina atrophy is a common symptom in women after menopause owing to decreasing estrogen levels. The most conventional treatment for this condition is estrogen cream. The shortcoming is its weak adhesion to the vagina mucus, thus requiring frequent daily application. In this study, BDDE was selected to crosslink and graft chitosan with thioglycolic acid, to form thiolated chitosan (CT) and improve the mucoadhesive properties of chitosan. Genistein was selected as the bioactive molecule that could exhibit estrogen-like properties for long-term treatment of vaginal atrophy. The efficacies of the materials were characterized and evaluated both in vitro and in vivo. Results showed that the mucoadhesive property of CT was approximately two-fold stronger against the constant flow than unmodified chitosan. CT with genistein (CT-G) was administered intravaginally every three days in vivo. It showed that the developed CT-G recover 54 % of the epithelium thickness of an atrophic vagina and ease vaginal atrophy.
Assuntos
Vaginite Atrófica/tratamento farmacológico , Quitosana/química , Genisteína/uso terapêutico , Hidrogéis/química , Tioglicolatos/química , Animais , Vaginite Atrófica/patologia , Células CACO-2 , Adesão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Feminino , Genisteína/química , Genisteína/metabolismo , Genisteína/farmacologia , Humanos , Hidrogéis/síntese química , Hidrogéis/farmacologia , Camundongos , Ratos , Ratos Sprague-Dawley , Vagina/patologiaRESUMO
B cell-mediated antibody response plays critical roles in protective immunity, as well as in the pathogenesis of allergic and autoimmune diseases. Epigenetic histone and DNA modifications regulate gene transcription and immunity; however, so far, little is known about the role of epigenetic regulation in antibody responses. In this study, we found that mice deficient in the histone H2A deubiquitinase MYSM1, despite their severe defect in B cell development, exhibit an enhanced antibody response against both T cell-dependent and independent antigens. We revealed that MYSM1 intrinsically represses plasma cell differentiation and antibody production. Mechanistic studies demonstrated that MYSM1 is a transcriptional activator of Pax5, the repressors of plasma cell differentiation, by facilitating key transcriptional factor recruitment and coordinating histone modifications at the Pax5 loci. Hence, this study uncovers a critical role for MYSM1 in epigenetically repressing plasma cell differentiation and antibody production, in addition to its opposing, active role in B cell development. Importantly, this study further provides a new target and strategy to modulate antibody production and responses with profound therapeutic implications.
Assuntos
Formação de Anticorpos/genética , Endopeptidases/genética , Epigênese Genética , Regulação da Expressão Gênica , Animais , Linfócitos B/citologia , Linfócitos B/imunologia , Linfócitos B/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Proteínas de Ligação a DNA/genética , Imunoglobulinas/sangue , Imunoglobulinas/imunologia , Memória Imunológica , Ativação Linfocitária/imunologia , Camundongos , Camundongos Knockout , Fator de Transcrição PAX5/genética , Plasmócitos/citologia , Plasmócitos/imunologia , Plasmócitos/metabolismo , Fator 1 de Ligação ao Domínio I Regulador Positivo , Ligação Proteica , Proteínas Proto-Oncogênicas/metabolismo , Fatores de Transcrição de Fator Regulador X , Transativadores/metabolismo , Fatores de Transcrição/genética , Proteases Específicas de UbiquitinaRESUMO
Histone modifications play critical roles in regulating immunity; however, little is known about the epigenetic control of natural killer (NK) cell development. Here, we found that NK cell development is severely impaired in mice deficient in the histone H2A deubiquitinase MYSM1. We demonstrated that MYSM1 is required for NK cell maturation but not for NK lineage specification and commitment. We also found that MYSM1 intrinsically controls this NK cell maturation. Mechanistic studies revealed that the expression of transcription factor, inhibitor of DNA-binding protein (ID2), a critical factor for NK cell development, is impaired in Mysm1(-/-) NK cells. MYSM1 interacts with nuclear factor IL-3 (NFIL3, also known as E4BP4), a critical factor for mouse NK cell development, and the recruitment of nuclear factor Il-3 to the ID2 locus is dependent on MYSM1. Further, we observed that MYSM1 is involved in maintaining an active chromatin at the ID2 locus to promote NK cell development. Hence this study demonstrates the critical epigenetic regulation of NK cell development by the histone H2A deubiquitinase MYSM1 through the transcriptional control of transcription factors important for NK cell development.
Assuntos
Imunidade Adaptativa/imunologia , Endopeptidases/imunologia , Epigênese Genética/imunologia , Células Matadoras Naturais/imunologia , Animais , Imunoprecipitação da Cromatina , Endopeptidases/genética , Endopeptidases/metabolismo , Citometria de Fluxo , Células HEK293 , Humanos , Immunoblotting , Proteína 2 Inibidora de Diferenciação/metabolismo , Interleucina-3/metabolismo , Células Matadoras Naturais/citologia , Camundongos , Camundongos Knockout , Reação em Cadeia da Polimerase em Tempo Real , Transativadores , Transdução Genética , Proteases Específicas de UbiquitinaRESUMO
Epigenetic histone modifications play critical roles in the control of gene transcription. Recently, an increasing number of histone H2A deubiquitinases have been identified and characterized. However, the physiological functions for this entire group of histone H2A deubiquitinases remain unknown. In this study, we revealed that the histone H2A deubiquitinase MYSM1 plays an essential and intrinsic role in early B cell development. MYSM1 deficiency results in a block in early B cell commitment and a defect of B cell progenitors in expression of EBF1 and other B lymphoid genes. We further demonstrated that MYSM1 derepresses EBF1 transcription in B cell progenitors by orchestrating histone modifications and transcription factor recruitment to the EBF1 locus. Thus, this study not only uncovers the essential role for MYSM1 in gene transcription during early B cell development but also underscores the biological significance of reversible epigenetic histone H2A ubiquitination.