Hedgehog Pathway Activation Requires Coreceptor-Catalyzed, Lipid-Dependent Relay of the Sonic Hedgehog Ligand

Wierbowski, Bradley M.; Petrov, Kostadin; Aravena, Laura; Gu, Garrick H.; Xu, Yangqing; Salic, Adrian

doi:10.1016/j.devcel.2020.09.017

Cited by 53 publications

(52 citation statements)

References 101 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The physiological relevance of such structures, therefore, depends on the question whether Disp and Scube2 -that both act upstream of Ptc -release Shh in unprocessed or in delipidated processed form. Support for the former possibility comes from three previous Shh overexpression studies (Creanga et al, 2012;Tukachinsky et al, 2012;Wierbowski et al, 2020), while this and previous work of our group demonstrate Shh release in delipidated processed form (Dierker et al, 2009;Jakobs et al, 2014;Jakobs et al, 2017;Ohlig et al, 2011;Ohlig et al, 2012;Schürmann et al, 2018). We explain these different findings by our Shh/Hhat coexpression system to generate physiologically relevant, dual-membrane associated Shh in nearquantitative manner.…”

Section: Discussionsupporting

confidence: 51%

“…As described before, this is because sheddases require membrane-proximal positioning of substrate cleavage sites for their activity, which is usually achieved by their membrane-association (Lichtenthaler, Lemberg, & Fluhrer, 2018). We also note that the current concept of lipidated Hh hand-over from Disp to Scube2 to GAS1 to Ptc (Creanga et al, 2012;Tukachinsky et al, 2012;Wierbowski et al, 2020) is in conflict with the in vivo observation that murine Disp expression in Disp-deficient Drosophila restores Hh dependent development (Burke et al, 1999), despite the lack of Scube orthologs in flies. It is also difficult to reconcile with the in vivo observation that transgenes with a 27kDa GFP tag inserted between the 19kDa Hh and its C-terminal cholesterol restore the development of Hh-deficient flies to adulthood (Chen, Huang, Hatori, & Kornberg, 2017), because this would require rather flexible mechanisms of Hh membrane extraction, repeated hand-over and receptor activation.…”

Section: Discussionmentioning

confidence: 80%

“…The resulting stable Scube2-Shh complex may then relay and signal to receiving cells that express the receptor Ptc. This model has been supported in vitro by Disp-and Scube2 co-immunoprecipitation with Shh (Creanga et al, 2012;Tukachinsky, Kuzmickas, Jao, Liu, & Salic, 2012), and recently published Scube2-Shh interactions with Ptc coreceptors Cdon/Boc and Gas1 (Wierbowski et al, 2020).…”

Section: Introductionmentioning

confidence: 74%

See 2 more Smart Citations

Conserved cholesterol-related activities of Dispatched drive Sonic hedgehog shedding from the cell membrane

Ehring

Manikowski

Froese

et al. 2020

Preprint

View full text Add to dashboard Cite

The Sonic hedgehog (Shh) pathway controls embryonic development and tissue homeostasis after birth. Long-lasting questions about this pathway are how dual-lipidated, firmly plasma membrane-associated Shh ligand is released from producing cells to signal to distant target cells, and how the resistance-nodulation-division transporter Dispatched (Disp) regulates this process. Here we show that Disp inactivation in Shh expressing cells specifically impairs proteolytic Shh release from its lipidated terminal peptides, a process called ectodomain shedding. Shh shedding from Disp-deficient cells was restored by pharmacological membrane cholesterol extraction and by overexpressed transgenic Disp or structurally related Patched (Ptc, a putative cholesterol transporter). These data suggest that Disp regulates physiological Shh function via controlled cell surface shedding and that molecular mechanisms shared by Disp and Ptc exercise such sheddase control.

show abstract

Section: Discussionsupporting

confidence: 51%

Section: Discussionmentioning

confidence: 80%

Section: Introductionmentioning

confidence: 74%

See 1 more Smart Citation

Conserved cholesterol-related activities of Dispatched drive Sonic hedgehog shedding from the cell membrane

Ehring

Manikowski

Froese

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…Biologically active SHH ligand secreted from the epithelium induced pathway activity in the adjacent mesenchyme with maximal induction of downstream Shh target genes occurring nearest the epithelial signaling source, mimicking in vivo gene expression gradients. Thus, this MPM, along with other recently developed 3D in vitro organoid models (Cederquist et al, 2019 ), may aid in our understanding of how concentration gradients are formed and, more specifically, address persistent questions of how secreted SHH ligand is shuttled through extracellular spaces while remaining available to bind transmembrane receptors to initiate signal transduction (Wierbowski et al, 2020 ). A novel organoid model consisting of human progenitor/stem cells designed to model disruption of palatal fusion, which occurs later in development was also described recently (Belair et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

A Microphysiological Approach to Evaluate Effectors of Intercellular Hedgehog Signaling in Development

Johnson

Vitek

Morgan

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Paracrine signaling in the tissue microenvironment is a central mediator of morphogenesis, and modeling this dynamic intercellular activity in vitro is critical to understanding normal and abnormal development. For example, Sonic Hedgehog (Shh) signaling is a conserved mechanism involved in multiple developmental processes and strongly linked to human birth defects including orofacial clefts of the lip and palate. SHH ligand produced, processed, and secreted from the epithelial ectoderm is shuttled through the extracellular matrix where it binds mesenchymal receptors, establishing a gradient of transcriptional response that drives orofacial morphogenesis. In humans, complex interactions of genetic predispositions and environmental insults acting on diverse molecular targets are thought to underlie orofacial cleft etiology. Consequently, there is a need for tractable in vitro approaches that model this complex cellular and environmental interplay and are sensitive to disruption across the multistep signaling cascade. We developed a microplate-based device that supports an epithelium directly overlaid onto an extracellular matrix-embedded mesenchyme, mimicking the basic tissue architecture of developing orofacial tissues. SHH ligand produced from the epithelium generated a gradient of SHH-driven transcription in the adjacent mesenchyme, recapitulating the gradient of pathway activity observed in vivo. Shh pathway activation was antagonized by small molecule inhibitors of epithelial secretory, extracellular matrix transport, and mesenchymal sensing targets, supporting the use of this approach in high-content chemical screening of the complete Shh pathway. Together, these findings demonstrate a novel and practical microphysiological model with broad utility for investigating epithelial-mesenchymal interactions and environmental signaling disruptions in development.

show abstract

“…While PTCH1 function is sufficient for SMO inhibition, HH signal reception also requires at least one of three coreceptors (CDON, BOC, GAS1). CDON, BOC, and GAS1 have overlapping roles and are collectively required for HH signaling (Allen et al , 2011; Izzi et al , 2011; Wierbowski et al , 2020; Zhang et al , 2011). Mice with targeted mutations in any one of these coreceptors have a selective and partial loss of HH pathway function.…”

Section: Introductionmentioning

confidence: 99%

Δ9-tetrahydrocannabinol inhibits Hedgehog-dependent patterning during development

Hong

Szutorisz

et al. 2021

Preprint

View full text Add to dashboard Cite

Many birth defects are thought to arise from a multifactorial etiology; i.e., interaction between genetic and environmental risk factors. The Hedgehog (HH) signaling pathway regulates myriad developmental processes, and pathway inhibition is associated with birth defects, including holoprosencephaly (HPE). Cannabinoids are HH pathway inhibitors, but little is known of their effects on HH-dependent processes in mammalian embryos, and their mechanism of action is unclear. We report here that the psychoactive cannabinoid Δ9-tetrahydrocannabinol (THC) induces two hallmark HH loss-of-function phenotypes (HPE and ventral neural tube patterning defects) in Cdon mutant mice, which have a subthreshold deficit in HH signaling. THC therefore acts as a “conditional teratogen”, dependent on a complementing but insufficient genetic insult. In vitro findings indicate that THC is a direct, albeit relatively weak, inhibitor of the essential HH pathway component, Smoothened. In contrast, the canonical THC receptor, cannabinoid receptor-type 1, is not required for THC to inhibit HH signaling. Cannabis consumption during pregnancy may contribute to the combination of risk factors underlying specific developmental disorders. These findings therefore have significant public health relevance.

show abstract

Hedgehog Pathway Activation Requires Coreceptor-Catalyzed, Lipid-Dependent Relay of the Sonic Hedgehog Ligand

Cited by 53 publications

References 101 publications

Conserved cholesterol-related activities of Dispatched drive Sonic hedgehog shedding from the cell membrane

Conserved cholesterol-related activities of Dispatched drive Sonic hedgehog shedding from the cell membrane

A Microphysiological Approach to Evaluate Effectors of Intercellular Hedgehog Signaling in Development

Δ9-tetrahydrocannabinol inhibits Hedgehog-dependent patterning during development

Contact Info

Product

Resources

About