RESUMO
Hutchinson-Gilford progeria syndrome (HGPS) is a rare, premature ageing syndrome in children. HGPS is normally caused by a mutation in the LMNA gene, encoding nuclear lamin A. The classical mutation in HGPS leads to the production of a toxic truncated version of lamin A, progerin, which retains a farnesyl group. Farnesyltransferase inhibitors (FTI), pravastatin and zoledronic acid have been used in clinical trials to target the mevalonate pathway in HGPS patients to inhibit farnesylation of progerin, in order to reduce its toxicity. Some other compounds that have been suggested as treatments include rapamycin, IGF1 and N-acetyl cysteine (NAC). We have analysed the distribution of prelamin A, lamin A, lamin A/C, progerin, lamin B1 and B2 in nuclei of HGPS cells before and after treatments with these drugs, an FTI and a geranylgeranyltransferase inhibitor (GGTI) and FTI with pravastatin and zoledronic acid in combination. Confirming other studies prelamin A, lamin A, progerin and lamin B2 staining was different between control and HGPS fibroblasts. The drugs that reduced progerin staining were FTI, pravastatin, zoledronic acid and rapamycin. However, drugs affecting the mevalonate pathway increased prelamin A, with only FTI reducing internal prelamin A foci. The distribution of lamin A in HGPS cells was improved with treatments of FTI, pravastatin and FTI + GGTI. All treatments reduced the number of cells displaying internal speckles of lamin A/C and lamin B2. Drugs targeting the mevalonate pathway worked best for progerin reduction, with zoledronic acid removing internal progerin speckles. Rapamycin and NAC, which impact on the MTOR pathway, both reduced both pools of progerin without increasing prelamin A in HGPS cell nuclei.
Assuntos
Lamina Tipo A/metabolismo , Ácido Mevalônico/metabolismo , Progéria/metabolismo , Linhagem Celular , Inibidores Enzimáticos/farmacologia , Humanos , Progéria/patologiaRESUMO
Modeling across multiple scales is a current challenge in Systems Biology, especially when applied to multicellular organisms. In this paper, we present an approach to model at different spatial scales, using the new concept of Hierarchically Colored Petri Nets (HCPN). We apply HCPN to model a tissue comprising multiple cells hexagonally packed in a honeycomb formation in order to describe the phenomenon of Planar Cell Polarity (PCP) signaling in Drosophila wing. We have constructed a family of related models, permitting different hypotheses to be explored regarding the mechanisms underlying PCP. In addition our models include the effect of well-studied genetic mutations. We have applied a set of analytical techniques including clustering and model checking over time series of primary and secondary data. Our models support the interpretation of biological observations reported in the literature.
Assuntos
Polaridade Celular/fisiologia , Drosophila/citologia , Modelos Biológicos , Biologia de Sistemas/métodos , Asas de Animais/citologia , Animais , Análise por Conglomerados , MutaçãoRESUMO
Hutchinson-Gilford Progeria Syndrome (HGPS) is a severe premature aging syndrome that affects children. These children display characteristics associated with normal aging and die young usually from cardiovascular problems or stroke. Classical HGPS is caused by mutations in the gene encoding the nuclear structural protein lamin A. This mutation leads to a novel version of lamin A that retains a farnesyl group from its processing. This protein is called Progerin and is toxic to cellular function. Pre-lamin A is an immature version of lamin A and also has a farnesylation modification, which is cleaved in the maturation process to create lamin A.
Assuntos
Pesquisa Biomédica , Mutação , Proteínas Nucleares/metabolismo , Progéria/metabolismo , Precursores de Proteínas/metabolismo , Universidades , Inglaterra , Humanos , Lamina Tipo A , Proteínas Nucleares/genética , Progéria/genética , Precursores de Proteínas/genéticaRESUMO
The laminopathy Hutchinson-Gilford progeria syndrome (HGPS) is caused by the mutant lamin A protein progerin and leads to premature aging of affected children. Despite numerous cell biological and biochemical insights into the basis for the cellular abnormalities seen in HGPS, the mechanism linking progerin to the organismal phenotype is not fully understood. To begin to address the mechanism behind HGPS using Drosophila melanogaster, we have ectopically expressed progerin and lamin A. We found that ectopic progerin and lamin A phenocopy several effects of laminopathies in developing and adult Drosophila, but that progerin causes a stronger phenotype than wild-type lamin A.
Assuntos
Drosophila melanogaster/metabolismo , Progéria/patologia , Animais , Núcleo Celular/metabolismo , Núcleo Celular/patologia , Modelos Animais de Doenças , Drosophila melanogaster/embriologia , Lamina Tipo A/metabolismo , LongevidadeRESUMO
Planar cell polarity (PCP) signaling generates subcellular asymmetry along an axis orthogonal to the epithelial apical-basal axis. Through a poorly understood mechanism, cell clones that have mutations in some PCP signaling components, including some, but not all, alleles of the receptor frizzled, cause polarity disruptions of neighboring wild-type cells, a phenomenon referred to as domineering nonautonomy. Here, a contact-dependent signaling hypothesis, derived from experimental results, is shown by reaction-diffusion, partial differential equation modeling and simulation to fully reproduce PCP phenotypes, including domineering nonautonomy, in the Drosophila wing. The sufficiency of this model and the experimental validation of model predictions reveal how specific protein-protein interactions produce autonomy or domineering nonautonomy.
Assuntos
Polaridade Celular , Drosophila/citologia , Modelos Biológicos , Transdução de Sinais , Asas de Animais/citologia , Proteínas Adaptadoras de Transdução de Sinal , Alelos , Animais , Membrana Celular/metabolismo , Difusão , Proteínas Desgrenhadas , Drosophila/genética , Drosophila/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Retroalimentação Fisiológica , Receptores Frizzled , Matemática , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mutação , Fenótipo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Ligação Proteica , Receptores Acoplados a Proteínas G , Asas de Animais/metabolismoRESUMO
Some epithelial cells are polarized along an axis orthogonal to their apical-basal axes. Recent studies in Drosophila lead to the view that three classes of signaling molecules govern the planar cell polarity (PCP) pathway. The first class, or module, functions across whole tissues, providing directional information to individual cells. The second module, apparently shared by all planar polarized tissues, and related to the canonical Wnt signaling pathway, interprets the directional signal to produce subcellular asymmetries. The third modules are tissue specific, acting to translate subcellular asymmetry into the appropriate morphological manifestations in the different cell types.
Assuntos
Drosophila/embriologia , Células Epiteliais/metabolismo , Transdução de Sinais , Animais , Padronização Corporal , Proteínas de Drosophila/metabolismo , Receptores Frizzled , Proteínas de Membrana/metabolismo , Modelos Biológicos , Mutação , Fenótipo , Células Fotorreceptoras de Invertebrados/embriologia , Receptores Acoplados a Proteínas G , Fatores de TempoRESUMO
Planar cell polarity signaling in Drosophila requires the receptor Frizzled and the cytoplasmic proteins Dishevelled and Prickle. From initial, symmetric subcellular distributions in pupal wing cells, Frizzled and Dishevelled become highly enriched at the distal portion of the cell cortex. We describe a Prickle-dependent intercellular feedback loop that generates asymmetric Frizzled and Dishevelled localization. In the absence of Prickle, Frizzled and Dishevelled remain symmetrically distributed. Prickle localizes to the proximal side of pupal wing cells and binds the Dishevelled DEP domain, inhibiting Dishevelled membrane localization and antagonizing Frizzled accumulation. This activity is linked to Frizzled activity on the adjacent cell surface. Prickle therefore functions in a feedback loop that amplifies differences between Frizzled levels on adjacent cell surfaces.