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Mark E Baratz - One of the best experts on this subject based on the ideXlab platform.
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Fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in Dupuytren's Contracture
BMC Medical Genomics, 2012Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, Anna Njarlangattil, Christina Raykha, John Michael Krill-burger, Phillip H Gallo, David B O'gorman, Bing Siang Gan, Mark E BaratzAbstract:Background Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. Methods To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. Results We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. Conclusions These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release.
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fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in dupuytren s contracture
BMC Medical Genomics, 2012Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, Anna Njarlangattil, Christina Raykha, Phillip H Gallo, Bing Siang Gan, John M Krillburger, David B Ogorman, Mark E BaratzAbstract:Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release.
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reversal of tgf β1 stimulation of α smooth muscle actin and extracellular matrix components by cyclic amp in dupuytren s derived fibroblasts
BMC Musculoskeletal Disorders, 2011Co-Authors: Latha Satish, Sandra Johnson, Phillip H Gallo, Mark E Baratz, Sandeep KathjuAbstract:Background: Myofibroblasts, a derived subset of fibroblasts especially important in scar formation and wound contraction, have been found at elevated levels in affected Dupuytren’s tissues. Transformation of fibroblasts to myofibroblasts is characterized by expression of alpha- smooth muscle actin (a-SMA) and increased production of extracellular matrix (ECM) components, both events of relevance to connective tissue remodeling. We propose that increasing the activation of the cyclic AMP (cAMP)/protein kinase A signaling pathway will inhibit transforming growth factor-beta1 (TGF-b1)-induced ECM synthesis and myofibroblast formation and may provide a means to blunt fibrosis. Methods: Fibroblasts derived from areas of Dupuytren’s contracture cord (DC), from adjacent and phenotypically normal Palmar Fascia (PF), and from Palmar Fascia from patients undergoing carpal tunnel release (CTR; CT) were treated with TGF-b1 (2 ng/ml) and/or forskolin (10 μM) (a known stimulator of cAMP). Total RNA and protein extracted was subjected to real time RT-PCR and Western blot analysis. Results: The basal mRNA expression levels of fibronectin- extra domain A (FN1-EDA), type I (COL1A2) and type III collagen (COL3A1), and connective tissue growth factor (CTGF) were all significantly increased in DC- and in PFderived cells compared to CT-derived fibroblasts. The TGF-b1 stimulation of a-SMA, CTGF, COL1A2 and COL3A1 was greatly inhibited by concomitant treatment with forskolin, especially in DC-derived cells. In contrast, TGF-b1 stimulation of FN1-EDA showed similar levels of reduction with the addition of forskolin in all three cell types. Conclusion: In sum, increasing cAMP levels show potential to inhibit the formation of myofibroblasts and accumulation of ECM components. Molecular agents that increase cAMP may therefore prove useful in mitigating DC progression or recurrence.
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Identification of differentially expressed genes in fibroblasts derived from patients with Dupuytren's Contracture
BMC Medical Genomics, 2008Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, David B O'gorman, Bing Siang Gan, Mark E Baratz, Benjamin Janto, J Christopher Post, Garth D Ehrlich, Sandeep KathjuAbstract:Dupuytren's contracture (DC) is the most common inherited connective tissue disease of humans and is hypothesized to be associated with aberrant wound healing of the Palmar Fascia. Fibroblasts and myofibroblasts are believed to play an important role in the genesis of DC and the fibroproliferation and contraction that are hallmarks of this disease. This study compares the gene expression profiles of fibroblasts isolated from DC patients and controls in an attempt to identify key genes whose regulation might be significantly altered in fibroblasts found within the Palmar Fascia of Dupuytren's patients. Total RNA isolated from diseased Palmar Fascia (DC) and normal Palmar Fascia (obtained during carpal tunnel release; 6 samples per group) was subjected to quantitative analyses using two different microarray platforms (GE Code Link™ and Illumina™) to identify and validate differentially expressed genes. The data obtained was analyzed using The Significance Analysis of Microarrays (SAM) software through which we identified 69 and 40 differentially regulated gene transcripts using the CodeLink™ and Illumina™ platforms, respectively. The CodeLink™ platform identified 18 upregulated and 51 downregulated genes. Using the Illumina™ platform, 40 genes were identified as downregulated, eleven of which were identified by both platforms. Quantitative RT-PCR confirmed the downregulation of three high-interest candidate genes which are all components of the extracellular matrix: proteoglycan 4 (PRG4), fibulin-1 (FBLN-1) transcript variant D, and type XV collagen alpha 1 chain. Overall, our study has identified a variety of candidate genes that may be involved in the pathophysiology of Dupuytren's contracture and may ultimately serve as attractive molecular targets for alternative therapies.
Latha Satish - One of the best experts on this subject based on the ideXlab platform.
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Fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in Dupuytren's Contracture
BMC Medical Genomics, 2012Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, Anna Njarlangattil, Christina Raykha, John Michael Krill-burger, Phillip H Gallo, David B O'gorman, Bing Siang Gan, Mark E BaratzAbstract:Background Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. Methods To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. Results We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. Conclusions These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release.
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fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in dupuytren s contracture
BMC Medical Genomics, 2012Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, Anna Njarlangattil, Christina Raykha, Phillip H Gallo, Bing Siang Gan, John M Krillburger, David B Ogorman, Mark E BaratzAbstract:Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release.
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reversal of tgf β1 stimulation of α smooth muscle actin and extracellular matrix components by cyclic amp in dupuytren s derived fibroblasts
BMC Musculoskeletal Disorders, 2011Co-Authors: Latha Satish, Sandra Johnson, Phillip H Gallo, Mark E Baratz, Sandeep KathjuAbstract:Background: Myofibroblasts, a derived subset of fibroblasts especially important in scar formation and wound contraction, have been found at elevated levels in affected Dupuytren’s tissues. Transformation of fibroblasts to myofibroblasts is characterized by expression of alpha- smooth muscle actin (a-SMA) and increased production of extracellular matrix (ECM) components, both events of relevance to connective tissue remodeling. We propose that increasing the activation of the cyclic AMP (cAMP)/protein kinase A signaling pathway will inhibit transforming growth factor-beta1 (TGF-b1)-induced ECM synthesis and myofibroblast formation and may provide a means to blunt fibrosis. Methods: Fibroblasts derived from areas of Dupuytren’s contracture cord (DC), from adjacent and phenotypically normal Palmar Fascia (PF), and from Palmar Fascia from patients undergoing carpal tunnel release (CTR; CT) were treated with TGF-b1 (2 ng/ml) and/or forskolin (10 μM) (a known stimulator of cAMP). Total RNA and protein extracted was subjected to real time RT-PCR and Western blot analysis. Results: The basal mRNA expression levels of fibronectin- extra domain A (FN1-EDA), type I (COL1A2) and type III collagen (COL3A1), and connective tissue growth factor (CTGF) were all significantly increased in DC- and in PFderived cells compared to CT-derived fibroblasts. The TGF-b1 stimulation of a-SMA, CTGF, COL1A2 and COL3A1 was greatly inhibited by concomitant treatment with forskolin, especially in DC-derived cells. In contrast, TGF-b1 stimulation of FN1-EDA showed similar levels of reduction with the addition of forskolin in all three cell types. Conclusion: In sum, increasing cAMP levels show potential to inhibit the formation of myofibroblasts and accumulation of ECM components. Molecular agents that increase cAMP may therefore prove useful in mitigating DC progression or recurrence.
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Identification of differentially expressed genes in fibroblasts derived from patients with Dupuytren's Contracture
BMC Medical Genomics, 2008Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, David B O'gorman, Bing Siang Gan, Mark E Baratz, Benjamin Janto, J Christopher Post, Garth D Ehrlich, Sandeep KathjuAbstract:Dupuytren's contracture (DC) is the most common inherited connective tissue disease of humans and is hypothesized to be associated with aberrant wound healing of the Palmar Fascia. Fibroblasts and myofibroblasts are believed to play an important role in the genesis of DC and the fibroproliferation and contraction that are hallmarks of this disease. This study compares the gene expression profiles of fibroblasts isolated from DC patients and controls in an attempt to identify key genes whose regulation might be significantly altered in fibroblasts found within the Palmar Fascia of Dupuytren's patients. Total RNA isolated from diseased Palmar Fascia (DC) and normal Palmar Fascia (obtained during carpal tunnel release; 6 samples per group) was subjected to quantitative analyses using two different microarray platforms (GE Code Link™ and Illumina™) to identify and validate differentially expressed genes. The data obtained was analyzed using The Significance Analysis of Microarrays (SAM) software through which we identified 69 and 40 differentially regulated gene transcripts using the CodeLink™ and Illumina™ platforms, respectively. The CodeLink™ platform identified 18 upregulated and 51 downregulated genes. Using the Illumina™ platform, 40 genes were identified as downregulated, eleven of which were identified by both platforms. Quantitative RT-PCR confirmed the downregulation of three high-interest candidate genes which are all components of the extracellular matrix: proteoglycan 4 (PRG4), fibulin-1 (FBLN-1) transcript variant D, and type XV collagen alpha 1 chain. Overall, our study has identified a variety of candidate genes that may be involved in the pathophysiology of Dupuytren's contracture and may ultimately serve as attractive molecular targets for alternative therapies.
William A Laframboise - One of the best experts on this subject based on the ideXlab platform.
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Fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in Dupuytren's Contracture
BMC Medical Genomics, 2012Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, Anna Njarlangattil, Christina Raykha, John Michael Krill-burger, Phillip H Gallo, David B O'gorman, Bing Siang Gan, Mark E BaratzAbstract:Background Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. Methods To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. Results We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. Conclusions These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release.
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fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in dupuytren s contracture
BMC Medical Genomics, 2012Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, Anna Njarlangattil, Christina Raykha, Phillip H Gallo, Bing Siang Gan, John M Krillburger, David B Ogorman, Mark E BaratzAbstract:Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release.
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Fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in Dupuytren's Contracture
'Springer Science and Business Media LLC', 2012Co-Authors: Satish L, William A Laframboise, John Michael Krill-burger, Johnson S, Vi L, Njarlangattil A, Raykha C, Ph Gallo, Db O'gorman, Bs GanAbstract:Background: Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. Methods. To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. Results: We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. Conclusions: These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release. © 2012 Satish et al; licensee BioMed Central Ltd
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Identification of differentially expressed genes in fibroblasts derived from patients with Dupuytren's Contracture
BMC Medical Genomics, 2008Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, David B O'gorman, Bing Siang Gan, Mark E Baratz, Benjamin Janto, J Christopher Post, Garth D Ehrlich, Sandeep KathjuAbstract:Dupuytren's contracture (DC) is the most common inherited connective tissue disease of humans and is hypothesized to be associated with aberrant wound healing of the Palmar Fascia. Fibroblasts and myofibroblasts are believed to play an important role in the genesis of DC and the fibroproliferation and contraction that are hallmarks of this disease. This study compares the gene expression profiles of fibroblasts isolated from DC patients and controls in an attempt to identify key genes whose regulation might be significantly altered in fibroblasts found within the Palmar Fascia of Dupuytren's patients. Total RNA isolated from diseased Palmar Fascia (DC) and normal Palmar Fascia (obtained during carpal tunnel release; 6 samples per group) was subjected to quantitative analyses using two different microarray platforms (GE Code Link™ and Illumina™) to identify and validate differentially expressed genes. The data obtained was analyzed using The Significance Analysis of Microarrays (SAM) software through which we identified 69 and 40 differentially regulated gene transcripts using the CodeLink™ and Illumina™ platforms, respectively. The CodeLink™ platform identified 18 upregulated and 51 downregulated genes. Using the Illumina™ platform, 40 genes were identified as downregulated, eleven of which were identified by both platforms. Quantitative RT-PCR confirmed the downregulation of three high-interest candidate genes which are all components of the extracellular matrix: proteoglycan 4 (PRG4), fibulin-1 (FBLN-1) transcript variant D, and type XV collagen alpha 1 chain. Overall, our study has identified a variety of candidate genes that may be involved in the pathophysiology of Dupuytren's contracture and may ultimately serve as attractive molecular targets for alternative therapies.
Sandra Johnson - One of the best experts on this subject based on the ideXlab platform.
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Fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in Dupuytren's Contracture
BMC Medical Genomics, 2012Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, Anna Njarlangattil, Christina Raykha, John Michael Krill-burger, Phillip H Gallo, David B O'gorman, Bing Siang Gan, Mark E BaratzAbstract:Background Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. Methods To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. Results We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. Conclusions These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release.
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fibroblasts from phenotypically normal Palmar Fascia exhibit molecular profiles highly similar to fibroblasts from active disease in dupuytren s contracture
BMC Medical Genomics, 2012Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, Anna Njarlangattil, Christina Raykha, Phillip H Gallo, Bing Siang Gan, John M Krillburger, David B Ogorman, Mark E BaratzAbstract:Dupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the Palmar Fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected Palmar Fascia, adjacent phenotypically normal Palmar Fascia, and non-DC Palmar Fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC. To achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected Palmar Fascia, patient-matched unaffected Palmar Fascia, and Palmar Fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram. We found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the Palmar Fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected Palmar Fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder. These data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected Palmar Fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the Palmar Fascia of patients undergoing carpal tunnel release.
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reversal of tgf β1 stimulation of α smooth muscle actin and extracellular matrix components by cyclic amp in dupuytren s derived fibroblasts
BMC Musculoskeletal Disorders, 2011Co-Authors: Latha Satish, Sandra Johnson, Phillip H Gallo, Mark E Baratz, Sandeep KathjuAbstract:Background: Myofibroblasts, a derived subset of fibroblasts especially important in scar formation and wound contraction, have been found at elevated levels in affected Dupuytren’s tissues. Transformation of fibroblasts to myofibroblasts is characterized by expression of alpha- smooth muscle actin (a-SMA) and increased production of extracellular matrix (ECM) components, both events of relevance to connective tissue remodeling. We propose that increasing the activation of the cyclic AMP (cAMP)/protein kinase A signaling pathway will inhibit transforming growth factor-beta1 (TGF-b1)-induced ECM synthesis and myofibroblast formation and may provide a means to blunt fibrosis. Methods: Fibroblasts derived from areas of Dupuytren’s contracture cord (DC), from adjacent and phenotypically normal Palmar Fascia (PF), and from Palmar Fascia from patients undergoing carpal tunnel release (CTR; CT) were treated with TGF-b1 (2 ng/ml) and/or forskolin (10 μM) (a known stimulator of cAMP). Total RNA and protein extracted was subjected to real time RT-PCR and Western blot analysis. Results: The basal mRNA expression levels of fibronectin- extra domain A (FN1-EDA), type I (COL1A2) and type III collagen (COL3A1), and connective tissue growth factor (CTGF) were all significantly increased in DC- and in PFderived cells compared to CT-derived fibroblasts. The TGF-b1 stimulation of a-SMA, CTGF, COL1A2 and COL3A1 was greatly inhibited by concomitant treatment with forskolin, especially in DC-derived cells. In contrast, TGF-b1 stimulation of FN1-EDA showed similar levels of reduction with the addition of forskolin in all three cell types. Conclusion: In sum, increasing cAMP levels show potential to inhibit the formation of myofibroblasts and accumulation of ECM components. Molecular agents that increase cAMP may therefore prove useful in mitigating DC progression or recurrence.
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Identification of differentially expressed genes in fibroblasts derived from patients with Dupuytren's Contracture
BMC Medical Genomics, 2008Co-Authors: Latha Satish, William A Laframboise, Sandra Johnson, David B O'gorman, Bing Siang Gan, Mark E Baratz, Benjamin Janto, J Christopher Post, Garth D Ehrlich, Sandeep KathjuAbstract:Dupuytren's contracture (DC) is the most common inherited connective tissue disease of humans and is hypothesized to be associated with aberrant wound healing of the Palmar Fascia. Fibroblasts and myofibroblasts are believed to play an important role in the genesis of DC and the fibroproliferation and contraction that are hallmarks of this disease. This study compares the gene expression profiles of fibroblasts isolated from DC patients and controls in an attempt to identify key genes whose regulation might be significantly altered in fibroblasts found within the Palmar Fascia of Dupuytren's patients. Total RNA isolated from diseased Palmar Fascia (DC) and normal Palmar Fascia (obtained during carpal tunnel release; 6 samples per group) was subjected to quantitative analyses using two different microarray platforms (GE Code Link™ and Illumina™) to identify and validate differentially expressed genes. The data obtained was analyzed using The Significance Analysis of Microarrays (SAM) software through which we identified 69 and 40 differentially regulated gene transcripts using the CodeLink™ and Illumina™ platforms, respectively. The CodeLink™ platform identified 18 upregulated and 51 downregulated genes. Using the Illumina™ platform, 40 genes were identified as downregulated, eleven of which were identified by both platforms. Quantitative RT-PCR confirmed the downregulation of three high-interest candidate genes which are all components of the extracellular matrix: proteoglycan 4 (PRG4), fibulin-1 (FBLN-1) transcript variant D, and type XV collagen alpha 1 chain. Overall, our study has identified a variety of candidate genes that may be involved in the pathophysiology of Dupuytren's contracture and may ultimately serve as attractive molecular targets for alternative therapies.
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De terugkeer naar de werkvloer of het hernemen van de dagelijkse activiteiten na een behandeling voor de ziekte van Dupuytren (chirurgie versus collagenase clostridium histolyticum)
'Peeters Publishers', 2020Co-Authors: Degreef Ilse, Vermeulen Louise, Arnauts AnjaAbstract:Abstract Return to work or the return to the daily activities after Dupuytren treatment: surgery vs collagenase clostridium histolyticum Dupuytren’s contracture is a disease of the Palmar Fascia, leading to flexion contractures which affect the hand function. This study evaluates if patients are satisfied with their treatment. It also evaluates the amount of time to return to work after treatment. Patients, undergoing fasciectomy or collagenase injections in the University Hospital of Leuven between 1st of January 2013 and the 31st of December 2017, received an invitation letter for study participation at home. They were asked to fill in a questionnaire and an informed consent and to send both documents back. Afterwards, some extra data were extracted from the medical file to complete the documents. The response ratio was 48/144 in participants undergoing fasciectomy and 36/170 in participants undergoing collagenase injection. Tubiana staging was higher in the fasciectomy group. The mean time to return to work was 75 days (±158) after fasciectomy and 12 days (±19) after collagenase injection. Both groups showed a significant improvement in satisfaction.status: publishe
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De terugkeer naar de werkvloer of het hernemen van de dagelijkse activiteiten na een behandeling (chirurgie versus collagenase clostridium histolyticum) voor de ziekte van Dupuytren
'Peeters Publishers', 2019Co-Authors: Degreef Ilse, Vermeulen Louise, Arnauts AnjaAbstract:Abstract Return to work or the return to the daily activities after Dupuytren treatment: surgery vs collagenase clostridium histolyticum Dupuytren’s contracture is a disease of the Palmar Fascia, leading to flexion contractures which affect the hand function. This study evaluates if patients are satisfied with their treatment. It also evaluates the amount of time to return to work after treatment. Patients, undergoing fasciectomy or collagenase injections in the University Hospital of Leuven between 1st of January 2013 and the 31st of December 2017, received an invitation letter for study participation at home. They were asked to fill in a questionnaire and an informed consent and to send both documents back. Afterwards, some extra data were extracted from the medical file to complete the documents. The response ratio was 48/144 in participants undergoing fasciectomy and 36/170 in participants undergoing collagenase injection. Tubiana staging was higher in the fasciectomy group. The mean time to return to work was 75 days (±158) after fasciectomy and 12 days (±19) after collagenase injection. Both groups showedstatus: accepte
