Vertebrate Hedgehog is secreted on two types of extracellular vesicles with different signaling properties

Vyas, Neha; Walvekar, Ankita; Tate, Dhananjay; Lakshmanan, Vairavan; Bansal, Dhiru; Cicero, Alessandra Lo; Raposo, Graça; Palakodeti, Dasaradhi; Dhawan, Jyotsna

doi:10.1038/srep07357

Cited by 117 publications

(122 citation statements)

References 51 publications

(76 reference statements)

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“…Sonic Hedgehog (Shh) is a secreted morphogen carried by exosomes that elicits differentiation and patterning in developing tissues [21,22]. Shh is expressed in adult tissue and mediates cell differentiation.…”

Section: Introductionmentioning

confidence: 99%

Adipocyte-Derived Exosomes Carrying Sonic Hedgehog Mediate M1 Macrophage Polarization-Induced Insulin Resistance via Ptch and PI3K Pathways

Song

Han

Chen

et al. 2018

Cell Physiol Biochem

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Background/Aims: Adipocyte-derived exosomes (ADEs) stimulate the activation of macrophages and contribute to the development of insulin resistance. Sonic Hedgehog (Shh) is an exosome-carrying protein and stimulates macrophages to secrete inflammatory cytokines. However, the impact of ADEs carrying Shh on the pro-inflammatory activation of macrophages and consequently, adipocyte insulin resistance is unclear. Methods: 3T3-L1 adipocytes were cultured with high glucose and insulin to imitate the pathogeny of insulin resistance. ADEs were isolated from conditioned media of 3T3-L1 adipocytes via differential ultracentrifugation. We explored the role of ADEs carrying Shh in the polarization of macrophages by flow cytometry. Western blot and electrophoretic mobility shift assay (EMSA) were performed to determine the activation of Shh-mediated signalling pathways. The effects of ADE-treated macrophages on adipocyte insulin signalling were studied by Western blot. Results: We found that circulating Shh-positive exosomes were increased in type 2 diabetes patients. High glucose and insulin increased the secretion of Shh-positive ADEs. The ADEs carrying Shh induced pro-inflammatory or M1 polarization of bone marrow-derived macrophages (BMDM) and RAW 264.7 macrophages. Inhibitors of Ptch and PI3K blocked the M1 polarization induced by ADEs, which suggests that ADEs carrying Shh mediated M1 macrophage polarization through the Ptch/PI3K signalling pathway. ADE-treated RAW 264.7 macrophages were subsequently used to assess the effect on insulin signalling in adipocytes. Using a co-culture assay, we showed that both ADE-treated macrophages and exosomes from these macrophages could decrease the expression of insulin-resistant substrate-1 (IRS-1) and hormone-sensitive lipase (HSL) in adipocytes. Inhibitors of Ptch and PI3K blocked the down-regulation of IRS-1 and HSL induced by ADE-treated macrophages. Conclusion: Together, these data indicate that ADEs carrying Shh induce the M1 polarization of macrophages, which contributes to insulin resistance in adipocytes through the Ptch/PI3K pathway.

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Section: Introductionmentioning

confidence: 99%

Adipocyte-Derived Exosomes Carrying Sonic Hedgehog Mediate M1 Macrophage Polarization-Induced Insulin Resistance via Ptch and PI3K Pathways

Song

Han

Chen

et al. 2018

Cell Physiol Biochem

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“…Shh-mediated tissue patterning begins with the production and processing of the full-length precursor protein (Lee et al, 1994;Pepinsky et al, 1998;Porter et al, 1995Porter et al, , 1996, whereupon mature ShhNp is secreted in a short and long range manner through a number of mechanisms (Briscoe and Thérond, 2013;Callejo et al, 2011;D'Angelo et al, 2015;Gradilla et al, 2014;Guerrero and Kornberg, 2014;Matusek et al, 2014;Parchure et al, 2015;Vyas et al, 2014). In neurons, Shh utilizes the RSP as a mode of anterograde transport in axons (Beug et al, 2011); therefore, our identification of Sort1 from a GST-affinity screen provided a candidate sorting receptor for Shh.…”

Section: Discussionmentioning

confidence: 99%

Sortilin regulates sorting and secretion of Sonic hedgehog

et al. 2016

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Sonic Hedgehog (Shh) is a secreted morphogen that is an essential regulatorof patterning and growth. The Shh full-length protein undergoes autocleavage in the endoplasmic reticulum to generate the biologically active N-terminal fragment (ShhN), which is destined for secretion. We identified sortilin (Sort1), a member of the VPS10P-domain receptor family, as a new Shh trafficking receptor. We demonstrate that Sort-Shh interact by performing coimmunoprecipitation and proximity ligation assays in transfected cells and that they colocalize at the Golgi. Sort1 overexpression causes re-distribution of ShhN and, to a lesser extent, of full-length Shh to the Golgi and reduces Shh secretion. We show loss of Sort1 can partially rescue Hedgehog-associated patterning defects in a mouse model that is deficient in Shh processing, and we show that Sort1 levels negatively regulate anterograde Shh transport in axons in vitro and Hedgehog-dependent axon-glial interactions in vivo. Taken together, we conclude that Shh and Sort1 can interact at the level of the Golgi and that Sort1 directs Shh away from the pathways that promote its secretion.

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“…Infectious HCV particles are assembled in the cytoplasm of Huh-7.5 cells [13] before their association to lipoproteins [8,77] or exosomes [75,78] . The associations to lipoprotein particles and extracellular particles have been proposed as mechanisms to regulate vertebrate Hedgehog dispersion during development [79] . In addition, ApoE [25,80] and extracellular particles [47] are suspected to contribute also to the pathogenesis of viruses unrelated to HCV.…”

Section: Discussionmentioning

confidence: 99%

Assembly and release of infectious hepatitis C virus involving unusual organization of the secretory pathway

Triyatni

Berger

Saunier

2016

WJH

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Institutional review board statement: Approved the use of serum IgG of a patient cured from an HCV infection for in vitro studies (Hôpital and Institut Cochin, Paris).Institutional animal care and use committee statement: Not applicable.Conflict-of-interest statement: Saunier B, Triyatni M and Berger EA are co-inventors on NIH-owned patent #US 9052321 B2 on the HCV particle production system. Triyatni M et al . Unusual secretory pathway of hepatitis C virus in thin section transmission electron microscopy. These findings were compared with the JFH-1 strain/Huh-7.5 cell model. RESULTS:We found that CANX was necessary for the production of HCV particles by BHK-WNV cells. This process involved the recruitment of a subset of HCV proteins, detected by immunoglobulin of an HCV-cured patient, in a compartment of rearranged membranes bypassing the endoplasmic reticulum-Golgi intermediary compartment and surrounded by mitochondria. It also involved the maturation of N-linked glycans on HCV envelope proteins, which was required for assembly and/or secretion of HCV particles. The formation of this specialized compartment required RAB1; upon expression of HCV structural genes, this compartment developed large vesicles with viral particles. RAB1 and alpha-tubulin were required for the release of HCV particles. These cellular factors were also involved in the production of HCVcc in the JFH-1 strain/Huh-7.5 cell system, which involves HCV RNA replication. The secretion of HCV particles by BHK-WNV cells presents similarities with a pathway involving caspase-1; a caspase-1 inhibitor was found to suppress the production of HCV particles from a full-length genome. CONCLUSION:Prior activity of the WNV subgenomic replicon in BHK-21 cells promoted re-wiring of host factors for the assembly and release of infectious HCV in a caspase-1-dependent mechanism. Core tip: Our system for production of authentic infectious hepatitis C virus (HCV) in non-humanized, nonhepatic cells involves the rearrangement of inner cellular membranes triggered by the replication of flaviviruses. The present results suggest that this feature relies on the re-wiring of host factors that usually contribute to the secretion of glycoproteins to generate an unusual secretory pathway. This model offers a new way to study the properties of free HCV particles, i.e. , independently from lipoproteins.Triyatni M, Berger EA, Saunier B. Assembly and release of infectious hepatitis C virus involving unusual organization of the secretory pathway.

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Vertebrate Hedgehog is secreted on two types of extracellular vesicles with different signaling properties

Cited by 117 publications

References 51 publications

Adipocyte-Derived Exosomes Carrying Sonic Hedgehog Mediate M1 Macrophage Polarization-Induced Insulin Resistance via Ptch and PI3K Pathways

Adipocyte-Derived Exosomes Carrying Sonic Hedgehog Mediate M1 Macrophage Polarization-Induced Insulin Resistance via Ptch and PI3K Pathways

Sortilin regulates sorting and secretion of Sonic hedgehog

Assembly and release of infectious hepatitis C virus involving unusual organization of the secretory pathway

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