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A09 - Scaffolding of mechanotransduction domains in migratinfg and non-migrating cells

Principal Investigators

Prof. Dr. Gary R. Lewin, MDC

Dr. Annette Hammes-Lewin, MDC

The mechanosensitive ion channels, PIEZO1 and PIEZO2, are involved in multiple cellular processes, ranging from vessel formation to touch sensation (Ranade et al., 2014). We have previously shown that STOML3 modulates PIEZO channels and is present in oligomeric form in membrane micro domains (Wetzel et al., 2017, Poole et al., 2014). We also identified a range of stomatin‐domain modulating small molecules that either decrease or increase STOML3 domain‐size in the plasma membrane as measured with dSTORM super‐resolution microscopy (Wetzel et al., 2017). Recently, we made the surprising observation that >90% of PIEZO channels cannot be opened by mechanical stimuli under physiological membrane potentials (Moroni et al., 2018). However, very positive voltage steps or disease-causing mutations near the channel pore can release channels for efficient opening (Moroni et al., 2018). The same disease-causing mutations that change PIEZO2 voltage sensitivity are associated with craniofacial abnormalities in humans.

There is increasing evidence that mechanosensitive PIEZO proteins play a role in cell migration during development by sensing the mechanical environment during cell movement. We recently found that there is a very early migratory population of cells that express PIEZO2 channels that later in development contribute to the endothelial cell population in vessels throughout the mouse embryo. We will use transgenic mouse models to manipulate PIEZO sensitivity in these migrating cells for instance by miss‐expressing STOML3 in these cells. This will be done by using a Cre‐inducible overexpression of an epitope‐tagged STOML3 from the ROSA locus; STOML3 overexpression can initiated using a PIEZO2‐Cre mouse available to us (Ranade et al., 2014). We hypothesize that the scaffolding proteins like STOML3 may control the localization of mechanotransduction domains in migrating cells to sensitize mechanosensing regions of the cells, like filopodia, to mechanical perturbation. We also wish to label PIEZO2 expressing endothelial precursors to study the fine architecture of PIEZO protein localization in migrating cells.

We recently showed that both PIEZO1 and TRPV4 channels are required for fast mechanotransduction by chondrocytes in the joint; a stationary mechanosensitive cell type (Servin‐Vences et al., 2017). TRPV4 appears to be unique amongst TRP channels in that it is gated extremely efficiently in heterologous cells by substrate deflection (Servin‐Vences et al., 2017). We will use a TRPV1/TRPV4 chimeric channel approach to map the domain of TRPV4 that is required for substrate gating. We will use pull‐down approaches with proteomics to try and identify TRPV4‐interacting proteins that provide the mechanical link with the extracellular matrix. Such interacting proteins may represent novel scaffolding proteins with essential roles in mechanotransduction.

References:

  • Mecklenburg N, Kowalczyk I, Witte F, Görne J, Laier A, Mamo TM, Gonschior H, Lehmann M, Richter M, Sporbert A, Purfürst B, Hübner N, Hammes A. Identification of disease-relevant modulators of the SHH pathway in the developing brain. Development. 148(17):dev199307 (2021)

  • Kowalczyk I, Lee C, Schuster E, Hoeren J, Trivigno V, Riedel L, Görne J, Wallingford JB, Hammes A, Feistel K. Neural tube closure requires the endocytic receptor Lrp2 and its functional interaction with intracellular scaffolds. Development. 148(2):dev195008 (2021)

  • Moroni M, Servin-Vences MR, Fleischer R, Sánchez-Carranza O, Lewin GR. Voltage gating of mechanosensitive PIEZO channels. Nat Commun. 9(1):1096 (2018)
  • Servin-Vences MR, Moroni M, Lewin GR, Poole K. Direct measurement of TRPV4 and PIEZO1 activity reveals multiple mechanotransduction pathways in chondrocytes. Elife 6:e21074 (2017)
  • Wetzel C, Pifferi S, Picci C, Gök C, Hoffmann D, Bali KK, Lampe A, Lapatsina L, Fleischer R, Smith ES, Bégay V, Moroni M, Estebanez L, Kühnemund J, Walcher J, Specker E, Neuenschwander M, von Kries JP, Haucke V, Kuner R, Poulet JF, Schmoranzer J, Poole K, Lewin GR. Small-molecule inhibition of STOML3 oligomerization reverses pathological mechanical hypersensitivity. Nat Neurosci. 20(2):209-218 (2017)
  • Eccles RL, Czajkowski MT, Barth C, Müller PM, McShane E, Grunwald S, Beaudette P, Mecklenburg N, Volkmer R, Zühlke K, Dittmar G, Selbach M, Hammes A, Daumke O, Klussmann E, Urbé S, Rocks O. Bimodal antagonism of PKA signalling by ARHGAP36. Nat Commun. 7:12963 (2016)
  • Christ A, Christa A, Klippert J, Eule JC, Bachmann S, Wallace VA, Hammes A, Willnow TE. LRP2 Acts as SHH Clearance Receptor to Protect the Retinal Margin from Mitogenic Stimuli. Dev Cell. 35(1):36-48 (2015)
  • Frank JA, Moroni M, Moshourab R, Sumser M, Lewin GR, Trauner D. Photoswitchable fatty acids enable optical control of TRPV1. Nat Commun. 6:7118 (2015)
  • Schrenk-Siemens K, Wende H, Prato V, Song K, Rostock C, Loewer A, Utikal J, Lewin GR, Lechner SG, Siemens J. PIEZO2 is required for mechanotransduction in human stem cell-derived touch receptors. Nat Neurosci. 18(1):10-6 (2015)
  • Ranade SS, Woo SH, Dubin AE, Moshourab RA, Wetzel C, Petrus M, Mathur J, Bégay V, Coste B, Mainquist J, Wilson AJ, Francisco AG, Reddy K, Qiu Z, Wood JN, Lewin GR, Patapoutian A. Piezo2 is the major transducer of mechanical forces for touch sensation in mice. Nature. 516(7529):121-5 (2014)
  • Kur E, Mecklenburg N, Cabrera RM, Willnow TE, Hammes A. LRP2 mediates folate uptake in the developing neural tube. J Cell Sci. 127(Pt 10):2261-8 (2014)
  • Poole K, Herget R, Lapatsina L, Ngo HD, Lewin GR. Tuning Piezo ion channels to detect molecular-scale movements relevant for fine touch. Nat Commun. 5:3520 (2014)
  • Christ A, Christa A, Kur E, Lioubinski O, Bachmann S, Willnow TE, Hammes A. LRP2 is an auxiliary SHH receptor required to condition the forebrain ventral midline for inductive signals. Dev Cell. 22(2):268-78 (2012)
  • L. Lapatsina, J.A. Jira, E. St. J. Smith, K. Poole, A. Kozlenkov, D. Bilbao, G.R. Lewin and P.A. Heppenstall Regulation of ASIC channels by a stomatin/STOML3 complex located in a mobile vesicle pool in sensory neurons. Open Biology doi:10.1098/rsob.120096 (2012)

  • J. Brand, E. St J. Smith, D. Schwefel, L. Lapatsina, K. Poole, D. Omerbašić, A. Kozlenkov, J. Behlke, G.R. Lewin and O. Daumke. A Stomatin Dimer Modulates the Activity of Acid-sensing Ion Channels. The EMBO Journal 31: 3635–3646 (2012)

  • L.Y. Chiang, K. Poole, B.E. Oliveira, N. Duarte, Y.A. Bernal Sierra, L. Bruckner-Tuderman, M. Koch, J. Hu& G.R. Lewin. Laminin-332 coordinates mechanotransduction and growth cone bifurcation in sensory neurons.  Nat. Neurosci. 14, 993–1000 (2011)

  • L. Lapatsina, J. Brand, K. Poole, O. Daumke & G.R. Lewin. Stomatin-domain proteins. Eur. J. Cell Biol. 91, 240–245 (2011)

  • Gajera CR, Emich H, Lioubinski O, Christ A, Beckervordersandforth-Bonk R, Yoshikawa K, Bachmann S, Christensen EI, Götz M, Kempermann G, Peterson AS, Willnow TE, Hammes A. LRP2 in ependymal cells regulates BMP signaling in the adult neurogenic niche. J Cell Sci. 123(Pt 11):1922-30 (2010)
  • C. Wetzel, J. Hu, D. Riethmacher, A. Benckendorff, L. Harder, A. Eilers, R. Moshourab, A. Kozlenkov, D. Labuz, O. Caspani, B. Erdmann, H. Machelska, P.A. Heppenstall and G.R. Lewin. A stomatin-domain protein essential for touch sensation in the mouse. Nature 445, 206–9 (2007)