Evaluación de los niveles de expresión de n-acetilgalactosamina-6-sulfato sulfatasa y reducción de glicosaminoglicanos sobre fibroblastos msp iv y hek-293 empleando vectores virales

La mucopolisacaridosis IVA (MPS IVA) o síndrome de Morquio A pertenece al grupo de enfermedades de almacenamiento lisosomal. Esta enfermedad es causada por la deficiencia en la enzima lisosomal N-acetilgalactosamina-6-sulfato sulfatasa (GALNS), la cual degrada el querantán sulfato y condroitin-6-sulfato. Los pacientes con MPS IVA presentan características clínicas específicas tales como problemas respiratorios, apnea, enfermedades cardiacas, discapacidad visual, estatura baja, cuello corto, entre otros. Dentro de los posibles tratamientos para MPS IVA se encuentra la terapia génica. Avances recientes han demostrado que el uso de vectores AAV y lentivirales han generado niveles elevados de actividad GALNS. Sin embargo, para continuar con los estudios en terapia génica como un posible tratamiento para MPS IVA, se debe estudiar la correlación que existe entre la actividad de GALNS reportada después de la transducción con los efectos terapéuticos que se pueden presentar.Mucopolysaccharidosis tVA (MPS IV A), or Morquio A syndrome belongs to the group of lysosomal storage diseases. This disease is caused by a deficiency in the lysosomal enzyme Nacetylgalactosamine- 6-sulfate sulfatase (GALNS) that degrades keratan sulfate and chondroitin-6-sulfate. the patients with MPS IVA have specific clinical characteristics such as respiratory problems, sleep apne4 heart disease, visual impairment, short stature, short neck, rlmong others. Among the possible treafnents for MPS IVA is gene therapy. Recent advances have shown that the use of AAV vectors and lentiviral has generated high activity levels of GALNS. However, to continue studies in gene therapy as a possible treatment for MpS IVA, it should study the correlation between the activity of GALNS reported after transduction with therapeutic effects that may present.Biólogo (a)Pregrad

Bone Growth Induction in Mucopolysaccharidosis IVA Mouse

Mucopolysaccharidosis IVA (MPS IVA; Morquio A syndrome) is caused by a deficiency of the N-acetylgalactosamine-6-sulfate-sulfatase (GALNS) enzyme, leading to the accumulation of glycosaminoglycans (GAG), keratan sulfate (KS) and chondroitin-6-sulfate (C6S), mainly in cartilage and bone. This lysosomal storage disorder (LSD) is characterized by severe systemic skeletal dysplasia. To this date, none of the treatment options for the MPS IVA patients correct bone pathology. Enzyme replacement therapy with elosulfase alpha provides a limited impact on bone growth and skeletal lesions in MPS IVA patients. To improve bone pathology, we propose a novel gene therapy with a small peptide as a growth-promoting agent for MPS IVA. A small molecule in this peptide family has been found to exert biological actions over the cardiovascular system. This work shows that an AAV vector expressing a C-type natriuretic (CNP) peptide induces bone growth in the MPS IVA mouse model. Histopathological analysis showed the induction of chondrocyte proliferation. CNP peptide also changed the pattern of GAG levels in bone and liver. These results suggest the potential for CNP peptide to be used as a treatment in MPS IVA patients

Primary lung cancer cell culture from transthoracic needle biopsy samples

Artículo de investigación1-14Lung cancer is the leading cause of cancer death in the world. The high mortality rate of this pathology is directly related to its late detection, since its symptoms can be masked by other diseases of lower risk. Although in recent years the number of research related to this subject has increased, molecular mechanisms that trigger this disease remains poorly understood. Experimental models are therefore vital for use in research. Immortalized cell lines have inherent limitations. Explanted tumoral cells obtained by transthoracic needle biopsy can be a potential source of primary culture of human lung tumor cells. Tumor specimens from 14 patients suspected of primary or metastatic lung cancer were obtained by CT-guided transthoracic lung biopsy. Solid tumors were mechanically disaggregated under a stereoscope. Cells were cultured in Base C growth media supplemented with 5% fetal bovine serum in 24-well cell culture plates. Primary lung cancer cell culture was successfully cultured from 12 out of 14 patients. Once a confluent monolayer was obtained, cells were enzymatically harvested and passaged to Petri culture dishes. These primary cell cultures were characterized by cytogenetic tests and gene expression analysis of diagnostic markers. These primary cell cultures revealed chromosome rearrangements and changes in their chromosome complement typical of tumoral cells. Additionally, Fluorescence in situ hybridization analysis demonstrated that three cultures exhibited EGFR amplification. Finally, expression profiles of CK7, NAPSIN A, TTF1, and P63 genes allowed in some cases to confirm sample tumor phenotype. These results demonstrate that primary lung cancer cell culture is possible from percutaneous puncture and provides an important biological source to asses and investigate the molecular mechanisms of lung cancer

Epitelial mesenchymal transition : from the molecular to physiologic

La transición epitelio mesénquima (EMT) es un proceso compuesto de diferentes fases, donde una célula epitelial adquiere un fenotipo mesenquimal. Dentro de los cambios involucrados se encuentran: pérdida de la polaridad celular, adquisición de una capacidad migratoria, capacidad invasora, resistencia a la apoptosis y aumento en la producción de componentes de la matriz extracelular. Todos estos cambios ocurren como una consecuencia de la activación y represión de genes involucrados con rutas de señalización específicas relacionadas con este evento. La EMT está relacionada con procesos fisiológicos y patológicos como el cáncer. Consta de tres fases: una de células no migratorias, células premigratorias y células migratorias; cada una de ellas producto de diferentes señales intra o extracelulares, factores de transcripción (TGF-B, Snail, TWIST, Sox, Slug, ZEB1, entre otras) y proteínas involucradas (E-cadherina, integrina, vimentina, ocludinas y claudinas).Revisión1-10Transition mesenchymal epithelium (EMT) is a process composed of different phases where an epithelial cell acquires a mesenchymal phenotype. Among the changes involved are: loss of cellular polarity, acquisition of a migratory capacity, invasive capacity, resistance to apoptosis, and increase in the production of components of the extracellular matrix. All these changes occur as a consequence of the activation and repression of genes involved with specific signaling pathways related to this event. EMT is related to physiological and pathological processes such as cancer. It consists of three phases: A phase of non-migratory cells, pre-migratory cells and migratory cells; (TGF-B, Snail, TWIST, Sox, Slug, ZEB1 among others), and proteins involved (E-cadherin, integrin, vimentin, occludins and claudins)

Bone Growth Induction in Mucopolysaccharidosis IVA Mouse

Mucopolysaccharidosis IVA (MPS IVA; Morquio A syndrome) is caused by a deficiency of the N-acetylgalactosamine-6-sulfate-sulfatase (GALNS) enzyme, leading to the accumulation of glycosaminoglycans (GAG), keratan sulfate (KS) and chondroitin-6-sulfate (C6S), mainly in cartilage and bone. This lysosomal storage disorder (LSD) is characterized by severe systemic skeletal dysplasia. To this date, none of the treatment options for the MPS IVA patients correct bone pathology. Enzyme replacement therapy with elosulfase alpha provides a limited impact on bone growth and skeletal lesions in MPS IVA patients. To improve bone pathology, we propose a novel gene therapy with a small peptide as a growth-promoting agent for MPS IVA. A small molecule in this peptide family has been found to exert biological actions over the cardiovascular system. This work shows that an AAV vector expressing a C-type natriuretic (CNP) peptide induces bone growth in the MPS IVA mouse model. Histopathological analysis showed the induction of chondrocyte proliferation. CNP peptide also changed the pattern of GAG levels in bone and liver. These results suggest the potential for CNP peptide to be used as a treatment in MPS IVA patients

Mucopolysaccharidoses: Cellular Consequences of Glycosaminoglycans Accumulation and Potential Targets

Mucopolysaccharidoses (MPSs) constitute a heterogeneous group of lysosomal storage disorders characterized by the lysosomal accumulation of glycosaminoglycans (GAGs). Although lysosomal dysfunction is mainly affected, several cellular organelles such as mitochondria, endoplasmic reticulum, Golgi apparatus, and their related process are also impaired, leading to the activation of pathophysiological cascades. While supplying missing enzymes is the mainstream for the treatment of MPS, including enzyme replacement therapy (ERT), hematopoietic stem cell transplantation (HSCT), or gene therapy (GT), the use of modulators available to restore affected organelles for recovering cell homeostasis may be a simultaneous approach. This review summarizes the current knowledge about the cellular consequences of the lysosomal GAGs accumulation and discusses the use of potential modulators that can reestablish normal cell function beyond ERT-, HSCT-, or GT-based alternatives

Role of RUNX2 transcription factor in epithelial mesenchymal transition in non-small cell lung cancer lung cancer : epigenetic control of the RUNX2 P1 promoter

Q2Q1Artículo original1-13Lung cancer has a high mortality rate in men and women worldwide. Approximately 15% of diagnosed patients with this type of cancer do not exceed the 5-year survival rate. Unfortunately, diagnosis is established in advanced stages, where other tissues or organs can be affected. In recent years, lineage-specific transcription factors have been associated with a variety of cancers. One such transcription factor possibly regulating cancer is RUNX2, the master gene of early and late osteogenesis. In thyroid and prostate cancer, it has been reported that RUNX2 regulates expression of genes important in tumor cell migration and invasion. In this study, we report on RUNX2/p57 overexpression in 16 patients with primary non-small cell lung cancer and/or metastatic lung cancer associated with H3K27Ac at P1 gene promoter region. In some patients, H3K4Me3 enrichment was also detected, in addition to WDR5, MLL2, MLL4, and UTX enzyme recruitment, members of the COMPASS-LIKE complex. Moreover, transforming growth factor-b induced RUNX2/p57 overexpression and specific RUNX2 knockdown supported a role for RUNX2 in epithelial mesenchymal transition, which was demonstrated through loss of function assays in adenocarcinoma A549 lung cancer cell line. Furthermore, RUNX2 increased expression of epithelial mesenchymal transition genes VIMENTIN, TWIST1, and SNAIL1, which reflected increased migratory capacity in lung adenocarcinoma cells

Transición epitelio mesénquima: de lo molecular a lo fisiológico

Resumen La transición epitelio mesénquima (EMT) es un proceso compuesto de diferentes fases, donde una célula epitelial adquiere un fenotipo mesenquimal. Dentro de los cambios involucrados se encuentran: pérdida de la polaridad celular, adquisición de una capacidad migratoria, capacidad invasora, resistencia a la apoptosis y aumento en la producción de componentes de la matriz extracelular. Todos estos cambios ocurren como una consecuencia de la activación y represión de genes involucrados con rutas de señalización específicas relacionadas con este evento. La EMT está relacionada con procesos fisiológicos y patológicos como el cáncer. Consta de tres fases: una de células no migratorias, células premigratorias y células migratorias; cada una de ellas producto de diferentes señales intra o extracelulares, factores de transcripción (TGF-B, Snail, TWIST, Sox, Slug, ZEB1, entre otras) y proteínas involucradas (E-cadherina, integrina, vimentina, ocludinas y claudinas).