{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T19:58:57Z","timestamp":1768507137910,"version":"3.49.0"},"posted":{"date-parts":[[2023,7,13]]},"group-title":"Biochemistry","reference-count":59,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,7,13]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Aggregation of leukocyte cell-derived chemotaxin 2 (LECT2) causes ALECT2, a systemic amyloidosis that affects the kidney and liver. Homozygosity of the I40V LECT2 mutation is believed to be necessary but not sufficient for the disease. Previous studies established that LECT2 fibrillogenesis is greatly accelerated by loss of its single bound zinc ion and stirring or shaking. These forms of agitation are often used to facilitate protein aggregation, but they create heterogeneous shear conditions, including air-liquid interfaces that denature proteins, that are not present in the body. Here, we determined the extent to which a more physiological form of mechanical stress\u2014shear generated by fluid flow through a network of artery and capillary-sized channels\u2014drives LECT2 fibrillogenesis. To mimic blood flow through the human kidney, where LECT2 and other proteins form amyloid deposits, we developed a microfluidic device consisting of progressively branched channels narrowing from 5 mm to 20 \u03bcm in width. Flow shear was particularly pronounced at the branch points and in the smallest capillaries, and this induced LECT2 aggregation much more efficiently than conventional shaking methods. EM images suggested the resulting fibril structures were different in the two conditions. Importantly, results from the microfluidic device showed the first evidence that the I40V mutation accelerated fibril formation and increased both size and density of the aggregates. These findings suggest that kidney-like flow shear, in combination with zinc loss, acts in combination with the I40V mutation to trigger LECT2 amyloidogenesis. These microfluidic devices may be of general use for uncovering the mechanisms by which blood flow induces misfolding and amyloidosis of circulating proteins.<\/jats:p>","DOI":"10.1101\/2023.07.13.548788","type":"posted-content","created":{"date-parts":[[2023,7,14]],"date-time":"2023-07-14T00:50:17Z","timestamp":1689295817000},"source":"Crossref","is-referenced-by-count":0,"title":["Mimicking Kidney Flow Shear Efficiently Induces Aggregation of LECT2, a Protein Involved in Renal Amyloidosis"],"prefix":"10.64898","author":[{"given":"Jeung-Hoi","family":"Ha","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yikang","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Harsimranjit","family":"Sekhon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephan","family":"Wilkens","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dacheng","family":"Ren","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4387-9644","authenticated-orcid":false,"given":"Stewart N.","family":"Loh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023071801350836000_2023.07.13.548788v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/ki.2008.152"},{"key":"2023071801350836000_2023.07.13.548788v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/ki.2013.558"},{"key":"2023071801350836000_2023.07.13.548788v1.3","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1146\/annurev-pathol-052016-100200","article-title":"Amyloidosis: Insights from Proteomics","volume":"12","year":"2017","journal-title":"Annu Rev Pathol"},{"key":"2023071801350836000_2023.07.13.548788v1.4","doi-asserted-by":"publisher","DOI":"10.2215\/CJN.12551214"},{"key":"2023071801350836000_2023.07.13.548788v1.5","first-page":"266","article-title":"Mixed leukocyte cell-derived chemotaxin 2 and amyloid A renal amyloidosis in a Kazakh-German patient","volume":"10","year":"2017","journal-title":"Clin Kidney J"},{"key":"2023071801350836000_2023.07.13.548788v1.6","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1111\/cts.12469","article-title":"Leukocyte Cell-Derived Chemotaxin-2: It\u2019s Role in Pathophysiology and Future in Clinical Medicine","volume":"10","year":"2017","journal-title":"Clin Transl Sci"},{"key":"2023071801350836000_2023.07.13.548788v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.100446"},{"key":"2023071801350836000_2023.07.13.548788v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2019.06.001"},{"key":"2023071801350836000_2023.07.13.548788v1.9","doi-asserted-by":"publisher","DOI":"10.1093\/protein\/gzp059"},{"key":"2023071801350836000_2023.07.13.548788v1.10","first-page":"1","article-title":"Shear-induced aggregation of amyloid \u03b2 (1\u201340) in a parallel plate geometry","volume":"0","year":"2020","journal-title":"Journal of Biomolecular Structure and Dynamics"},{"key":"2023071801350836000_2023.07.13.548788v1.11","first-page":"225","volume-title":"Progress in Molecular Biology and Translational Science","volume":"166","year":"2019"},{"key":"2023071801350836000_2023.07.13.548788v1.12","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1218402110"},{"key":"2023071801350836000_2023.07.13.548788v1.13","doi-asserted-by":"crossref","first-page":"5020","DOI":"10.1039\/b914089a","article-title":"Shear-induced structure and mechanics of \u03b2-lactoglobulin amyloid fibrils","volume":"5","year":"2009","journal-title":"Soft Matter"},{"key":"2023071801350836000_2023.07.13.548788v1.14","doi-asserted-by":"publisher","DOI":"10.1021\/bm0505078"},{"key":"2023071801350836000_2023.07.13.548788v1.15","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1016\/j.bpj.2009.10.010","article-title":"Susceptibility of different proteins to flow-induced conformational changes monitored with Raman spectroscopy","volume":"98","year":"2010","journal-title":"Biophys J"},{"key":"2023071801350836000_2023.07.13.548788v1.16","doi-asserted-by":"crossref","first-page":"3461","DOI":"10.1039\/C5SM02916C","article-title":"Shear-induced amyloid fibrillization: the role of inertia","volume":"12","year":"2016","journal-title":"Soft Matter"},{"key":"2023071801350836000_2023.07.13.548788v1.17","doi-asserted-by":"publisher","DOI":"10.1021\/bi002555c"},{"key":"2023071801350836000_2023.07.13.548788v1.18","doi-asserted-by":"crossref","first-page":"10355","DOI":"10.1021\/acs.jpcb.1c05171","article-title":"Effects of Shear Rate and Protein Concentration on Amyloidogenesis via Interfacial Shear","volume":"125","year":"2021","journal-title":"J. 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