eprintid: 10055981 rev_number: 32 eprint_status: archive userid: 608 dir: disk0/10/05/59/81 datestamp: 2018-09-18 13:23:42 lastmod: 2022-01-07 23:38:12 status_changed: 2018-09-18 13:23:42 type: article metadata_visibility: show creators_name: Loaiza, S creators_name: Ferreira, SA creators_name: Chinn, TM creators_name: Kirby, A creators_name: Tsolaki, E creators_name: Dondi, C creators_name: Parzych, K creators_name: Strange, AP creators_name: Bozec, L creators_name: Bertazzo, S creators_name: Hedegaard, MAB creators_name: Gentleman, E creators_name: Auner, HW title: An engineered, quantifiable in vitro model for analysing the effect of proteostasis-targeting drugs on tissue physical properties ispublished: pub divisions: UCL divisions: B02 divisions: C10 divisions: D21 divisions: B04 divisions: C05 divisions: F43 divisions: F42 keywords: Atomic force microscopy, Cancer diagnosis and therapy, Proteasome, Proteostasis, Raman spectroscopy, VCP/p97 note: © 2018 The Authors. This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/). abstract: Cellular function depends on the maintenance of protein homeostasis (proteostasis) by regulated protein degradation. Chronic dysregulation of proteostasis is associated with neurodegenerative and age-related diseases, and drugs targeting components of the protein degradation apparatus are increasingly used in cancer therapies. However, as chronic imbalances rather than loss of function mediate their pathogenesis, research models that allow for the study of the complex effects of drugs on tissue properties in proteostasis-associated diseases are almost completely lacking. Here, to determine the functional effects of impaired proteostatic fine-tuning, we applied a combination of materials science characterisation techniques to a cell-derived, in vitro model of bone-like tissue formation in which we pharmacologically perturbed protein degradation. We show that low-level inhibition of VCP/p97 and the proteasome, two major components of the degradation machinery, have remarkably different effects on the bone-like material that human bone-marrow derived mesenchymal stromal cells (hMSC) form in vitro. Specifically, whilst proteasome inhibition mildly enhances tissue formation, Raman spectroscopic, atomic force microscopy-based indentation, and electron microscopy imaging reveal that VCP/p97 inhibition induces the formation of bone-like tissue that is softer, contains less protein, appears to have more crystalline mineral, and may involve aberrant micro- and ultra-structural tissue organisation. These observations contrast with findings from conventional osteogenic assays that failed to identify any effect on mineralisation. Taken together, these data suggest that mild proteostatic impairment in hMSC alters the bone-like material they form in ways that could explain some pathologies associated with VCP/p97-related diseases. They also demonstrate the utility of quantitative materials science approaches for tackling long-standing questions in biology and medicine, and could form the basis for preclinical drug testing platforms to develop therapies for diseases stemming from perturbed proteostasis or for cancer therapies targeting protein degradation. Our findings may also have important implications for the field of tissue engineering, as the manufacture of cell-derived biomaterial scaffolds may need to consider proteostasis to effectively replicate native tissues. date: 2018-11 date_type: published official_url: https://doi.org/10.1016/j.biomaterials.2018.08.041 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green article_type_text: Journal Article verified: verified_manual elements_id: 1581944 doi: 10.1016/j.biomaterials.2018.08.041 pii: S0142-9612(18)30596-9 lyricists_name: Bertazzo, Sergio lyricists_name: Bozec, Laurent lyricists_name: Strange, Adam lyricists_name: Tsolaki, Elena lyricists_id: SBERT93 lyricists_id: LBOZE75 lyricists_id: STRAN38 lyricists_id: EETSO10 actors_name: Bracey, Alan actors_id: ABBRA90 actors_role: owner full_text_status: public publication: Biomaterials volume: 183 pagerange: 102-113 event_location: Netherlands issn: 1878-5905 citation: Loaiza, S; Ferreira, SA; Chinn, TM; Kirby, A; Tsolaki, E; Dondi, C; Parzych, K; ... Auner, HW; + view all <#> Loaiza, S; Ferreira, SA; Chinn, TM; Kirby, A; Tsolaki, E; Dondi, C; Parzych, K; Strange, AP; Bozec, L; Bertazzo, S; Hedegaard, MAB; Gentleman, E; Auner, HW; - view fewer <#> (2018) An engineered, quantifiable in vitro model for analysing the effect of proteostasis-targeting drugs on tissue physical properties. Biomaterials , 183 pp. 102-113. 10.1016/j.biomaterials.2018.08.041 <https://doi.org/10.1016/j.biomaterials.2018.08.041>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10055981/1/1-s2.0-S0142961218305969-main.pdf document_url: https://discovery.ucl.ac.uk/id/eprint/10055981/8/Loaiza_1-s2.0-S0142961218305969-mmc1.pdf