scholarly journals Silencing Heat Shock Protein 47 (HSP47) in Fibrogenic Precision-Cut Lung Slices: A Surprising Lack of Effects on Fibrogenesis?

2021 ◽  
Vol 8 ◽  
Author(s):  
Mitchel J. R. Ruigrok ◽  
Khaled E. M. El Amasi ◽  
Diana J. Leeming ◽  
Jannie M. B. Sand ◽  
Henderik W. Frijlink ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Transforming Growth Factor ◽  
Therapeutic Potential ◽  
Scar Tissue ◽  
Transforming Growth Factor Β1 ◽  
Heat Shock Protein 47 ◽  
Collagen Secretion ◽  
Precision Cut Lung Slices ◽  
Interfering Rna

Idiopathic pulmonary fibrosis (IPF) is a chronic disease that is characterized by the excessive deposition of scar tissue in the lungs. As currently available treatments are unable to restore lung function in patients, there is an urgent medical need for more effective drugs. Developing such drugs, however, is challenging because IPF has a complex pathogenesis. Emerging evidence indicates that heat shock protein 47 (HSP47), which is encoded by the gene Serpinh1, may be a suitable therapeutic target as it is required for collagen synthesis. Pharmacological inhibition or knockdown of HSP47 could therefore be a promising approach to treat fibrosis. The objective of this study was to assess the therapeutic potential of Serpinh1-targeting small interfering RNA (siRNA) in fibrogenic precision-cut lung slices prepared from murine tissue. To enhance fibrogenesis, slices were cultured for up to 144 h with transforming growth factor β1. Self-deliverable siRNA was used to knockdown mRNA and protein expression, without affecting the viability and morphology of slices. After silencing HSP47, only the secretion of fibronectin was reduced while other aspects of fibrogenesis remained unaffected (e.g., myofibroblast differentiation as well as collagen secretion and deposition). These observations are surprising as others have shown that Serpinh1-targeting siRNA suppressed collagen deposition in animals. Further studies are therefore warranted to elucidate downstream effects on fibrosis upon silencing HSP47.

Treatment of Idiopathic Myelofibrosis Employing siRNA for Heat Shock Protein 47 (siRNA/HSP47) Encapsulated in Liposomes.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4646-4646
Author(s):  
Junichi Watanabe ◽  
Takuya Matsunaga ◽  
Kazuyuki Murase ◽  
Yasushi Sato ◽  
Taiko Nagaya ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Fibroblast Cell ◽  
Idiopathic Myelofibrosis ◽  
Hematopoietic Stem ◽  
Heat Shock Protein 47 ◽  
Collagen Secretion ◽  
Novel Approach ◽  
Murine Fibroblast ◽  
Chronic Myeloproliferative Disorder

Abstract Idiopathic myelofibrosis (IMF) is a chronic myeloproliferative disorder characterized by bone marrow (BM) fibrosis, splenomegaly, extramedurally hematopoiesis and anemia. At present, there is no curative treatment available for this disease except allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the high therapy-related mortality of allo-HSCT, even in reduced-intensity transplantation, could not be ignored. We report a novel approach for the treatment of IMF, utilizing siRNA for a collagen specific chaperon, heat shock protein 47 (HSP47) encapsulated in liposomes (lip-siRNA/HSP47). We first confirmed suppression of HSP47 in NIH3T3 murine fibroblast cell line and primary murine BM fibroblasts, and collagen secretion from these cells by siRNA/HSP47. We next treated thrombopoietin (TPO) transgenic mice exhibiting IMF like feature (Shimoda K, Harada M et al. Leuk Res29, 761–769, 2005; Am J Hematol82, 802–806, 2007). In these mice, treatment by intravenous injection of lip-siRNA/HSP47 resulted in histological resolution of myelofibrosis associated with apoptotic death of BM fibroblasts, and improvement of anemia and splenomegaly. Thus, lip-siRNA/HSP47 therapy may be a promising treatment for IMF.

Abdominal Insufflation Decreases Blood Loss without Worsening the Inflammatory Response: Implications for Prehospital Control of Internal Bleeding

2008 ◽  
Vol 74 (4) ◽  
pp. 297-301 ◽  
Author(s):  
George C. Velmahos ◽  
Konstantinos Spaniolas ◽  
Malek Tabbara ◽  
Michael Duggan ◽  
Yongqing Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Heat Shock ◽  
Blood Loss ◽  
Inflammatory Response ◽  
Heat Shock Protein ◽  
Heat Shock Protein 70 ◽  
Splenic Injury ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1

Abdominal insufflation (AI) by carbon dioxide has been shown to decrease the rate of bleeding in different swine models of abdominal organ injuries. With development of appropriate tools, AI could be used to control bleeding temporarily in the prehospital setting. Concerns have been raised about the inflammatory response to AI, which could damage organs at a later stage despite initial hemostasis. We hypothesized that AI controls bleeding without inducing an unfavorable inflammatory response. An experimental splenic injury was caused in 28 Yorkshire pigs, which were randomized to: 1) standard resuscitation (n = 14) with crystalloids to a mean arterial pressure of 60 mm Hg, or 2) standard resuscitation and AI (n = 14) to an abdominal pressure of 20 cmH2O. The experiment lasted for 30 minutes, and intra-abdominal blood loss was measured. Blood serum interleukin 1β (IL-1β), transforming growth factor β1, and lung tissue heat shock protein 70 gene expression were measured at 0, 15, and 30 minutes, as markers of the inflammatory response. All animals survived to the end of the experiment. Total blood loss was significantly less in the AI group compared with the other standard resuscitation animals (733 ± 76 vs 1094 ± 153 mL, P = 0.049). The pH at the end of the experiment was significantly lower in the AI group (7.28 ± 0.02 vs 7.44 ± 0.05, P < 0.01) but there was no difference in lactate levels (1.5 ± 0.4 vs 1.7 ± 0.3, P = 0.7). Similarly, there was no difference in IL-1β, transforming growth factor β1, or lung tissue heat shock protein 70 gene expression between the two groups at any time point, although there was a trend towards lower IL-1β levels in the AI group. Our conclusion is that AI reduces blood loss from splenic injury without a measurable effect on the early inflammatory response in a clinically relevant animal model.

Influence of exogenous growth factors on the synthesis and secretion of collagen types I and III by explants of normal and healing rabbit ligaments

1994 ◽  
Vol 72 (9-10) ◽  
pp. 403-409 ◽  
Author(s):  
Patricia G. Murphy ◽  
Barbara J. Loitz ◽  
Cyril B. Frank ◽  
David A. Hart
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Collagen Synthesis ◽  
Transforming Growth Factor ◽  
Scar Tissue ◽  
Transforming Growth Factor Β1 ◽  
Type I ◽  
Collagen Types ◽  
Collagen Secretion ◽  
Tgf Β1

In this investigation it has been demonstrated that specific growth factors are able to modify collagen secretion in explants from healing rabbit medial collateral ligaments. The addition of 2.5 ng transforming growth factor β1 (TGF-β1)/mL to 3-week-old scar explants resulted in an increase in the total amount of collagen secreted. Analysis of collagen types I and III individually revealed that the increase mediated by TGF-β1 was due primarily to an increase in collagen type I secretion. This led to a ratio of type I : type III that is closer to that found in normal ligament tissue. The addition of 100 ng insulin-like growth factor 2 (IGF-2) to explant cultures of 3-week-old scar tissue also led to an increase in the quantity of collagen secreted, but the increase was in both type I and III collagens. These effects were observed to a lesser degree in 6-week-old scar tissue, and by 12 weeks postinjury, minimal effects of the growth factors on collagen synthesis was detected. Neither growth factor influenced collagen secretion by normal ligament or synovium. In contrast, IGF-1 (100 ng/mL) or basic fibroblast growth factor (bFGF) (10 ng/mL) did not exert a detectable effect on collagen secretion by any of the normal or healing tissues. These results indicate that TGF-β1 and IGF-2 can modify the metabolic activity of cells in explants of healing ligaments early after injury and may enhance the repair process leading to improved function.Key words: ligament healing, transforming growth factor β1, insulin-like growth factors 1 and 2, basic fibroblast growth factor, collagen synthesis.

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