Identification of serum protein biomarkers in individuals with Niemann-Pick disease, type C1

Identification of serum protein biomarkers in individuals with Niemann-Pick disease, type C1 | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Identification of serum protein biomarkers in individuals with Niemann-Pick disease, type C1 Khushboo Singhal, Matthew T. Menold, Niamh X. Cawley, Kiersten Campbell, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8833559/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 14 You are reading this latest preprint version Abstract Background Niemann-Pick disease, type C1 (NPC1), is a rare, fatal, neurodegenerative lysosomal disorder caused by pathological variants in NPC1 . Defects in lysosomal cholesterol transport result in the accumulation of unesterified cholesterol within the endo-lysosomal compartments. Delayed diagnosis, limited treatment options, and phenotypic heterogeneity characterized by a broad range of signs/symptoms underscore the urgent need for effective biomarkers to facilitate diagnosis, monitor disease progression and assess therapeutic response. The goal of this study was to identify serum protein biomarkers for NPC1. Methods Proximal Extension Assays (PEA) were used to determine relative protein expression levels from 68 serum samples from NPC1 individuals and 20 age-appropriate control serum samples. Statistical models identified NPC1 disease-specific effects after adjusting for covariates. Selected proteins were orthogonally validated by ELISA and correlated with assessments of both disease severity (Age of Neurological Onset (ANO) and Annual Severity Increment Score (ASIS)) and disease burden (NPC Neurological Severity Score (NSS). Results Quantifiable data was obtained on 2888 proteins, revealing 186 increased (adjusted log 2 FC ≥ 1) and 286 decreased (adjusted log 2 FC ≤ -1) proteins with adj. p-value < 0.1 when comparing NPC1 individuals not being treated with miglustat versus control serum samples. Using orthogonal assays, we confirmed significant elevations for seven proteins: TREM2, AgRP, CCL18, Cathepsin L, GPNMB, NPY, and HSD17B14, and a significant decrease of BDNF. We further identified 100 proteins whose abundance levels were significantly altered towards normal by miglustat treatment. We found the 17-domain NPC NSS to be correlated with protein levels in the PEA data. Orthogonally validated data correlated with the age of neurological onset. We also identified 25 differentially abundant serum proteins in NPC1 baseline samples which are predominantly expressed in brain regions. Conclusions The statistical analysis pipeline developed in this study is flexible and scalable and supports application to high-dimensional proteomic datasets. This study identified and validated serum proteins with altered expression in individuals with NPC1, responded to miglustat therapy, and correlated with disease severity or burden. These proteins may have clinical utility as biomarkers and provide insights into cellular mechanisms contributing to NPC1 disease pathology. Trial Registrations: NCT00344331 (Registration on 2006-06-23) Niemann-Pick disease type C1 NPC1 Neurodegeneration Serum Protein Biomarkers Proximal Extension Assay Full Text Additional Declarations No competing interests reported. Supplementary Files AdditionalFiguresFDP.pptx AdditionalmaterialFDP.docx AdditionalTablesFDP.xlsx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 07 Mar, 2026 Reviews received at journal 01 Mar, 2026 Reviews received at journal 27 Feb, 2026 Reviews received at journal 26 Feb, 2026 Reviewers agreed at journal 21 Feb, 2026 Reviewers agreed at journal 21 Feb, 2026 Reviewers agreed at journal 20 Feb, 2026 Reviewers agreed at journal 19 Feb, 2026 Reviewers agreed at journal 19 Feb, 2026 Reviewers agreed at journal 19 Feb, 2026 Reviewers invited by journal 19 Feb, 2026 Editor assigned by journal 18 Feb, 2026 Submission checks completed at journal 10 Feb, 2026 First submitted to journal 09 Feb, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Defects in lysosomal cholesterol transport result in the accumulation of unesterified cholesterol within the endo-lysosomal compartments. Delayed diagnosis, limited treatment options, and phenotypic heterogeneity characterized by a broad range of signs/symptoms underscore the urgent need for effective biomarkers to facilitate diagnosis, monitor disease progression and assess therapeutic response. The goal of this study was to identify serum protein biomarkers for NPC1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eProximal Extension Assays (PEA) were used to determine relative protein expression levels from 68 serum samples from NPC1 individuals and 20 age-appropriate control serum samples. Statistical models identified NPC1 disease-specific effects after adjusting for covariates. Selected proteins were orthogonally validated by ELISA and correlated with assessments of both disease severity (Age of Neurological Onset (ANO) and Annual Severity Increment Score (ASIS)) and disease burden (NPC Neurological Severity Score (NSS).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eQuantifiable data was obtained on 2888 proteins, revealing 186 increased (adjusted log\u003csub\u003e2\u003c/sub\u003eFC ≥ 1) and 286 decreased (adjusted log\u003csub\u003e2\u003c/sub\u003eFC ≤ -1) proteins with adj. p-value \u0026lt; 0.1 when comparing NPC1 individuals not being treated with miglustat versus control serum samples. Using orthogonal assays, we confirmed significant elevations for seven proteins: TREM2, AgRP, CCL18, Cathepsin L, GPNMB, NPY, and HSD17B14, and a significant decrease of BDNF. We further identified 100 proteins whose abundance levels were significantly altered towards normal by miglustat treatment. We found the 17-domain NPC NSS to be correlated with protein levels in the PEA data. Orthogonally validated data correlated with the age of neurological onset. We also identified 25 differentially abundant serum proteins in NPC1 baseline samples which are predominantly expressed in brain regions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe statistical analysis pipeline developed in this study is flexible and scalable and supports application to high-dimensional proteomic datasets. This study identified and validated serum proteins with altered expression in individuals with NPC1, responded to miglustat therapy, and correlated with disease severity or burden. These proteins may have clinical utility as biomarkers and provide insights into cellular mechanisms contributing to NPC1 disease pathology.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTrial Registrations: NCT00344331 (Registration on 2006-06-23)\u003c/strong\u003e\u003c/p\u003e","manuscriptTitle":"Identification of serum protein biomarkers in individuals with Niemann-Pick disease, type C1","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-23 06:58:55","doi":"10.21203/rs.3.rs-8833559/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-03-08T02:26:18+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-01T22:39:17+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-27T22:34:49+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-26T12:34:38+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"49521758266684820438314883149180319571","date":"2026-02-21T17:00:34+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"203784700604817391764437598544501806092","date":"2026-02-21T10:49:10+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"146528727042134398422595504381316302119","date":"2026-02-20T05:42:51+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"51969384835062567701286384741415129172","date":"2026-02-19T22:23:29+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"112168652899049959946666308786423836265","date":"2026-02-19T13:40:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"114684426258342894670524104446619530164","date":"2026-02-19T10:54:39+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-19T05:46:37+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-18T14:14:45+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-10T16:14:59+00:00","index":"","fulltext":""},{"type":"submitted","content":"Biomarker Research","date":"2026-02-09T18:27:07+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"biomarker-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bmre","sideBox":"Learn more about [Biomarker Research](http://biomarkerres.biomedcentral.com)","snPcode":"40364","submissionUrl":"https://submission.nature.com/new-submission/40364/3","title":"Biomarker Research","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f8a8a3ac-cf92-4a18-8f24-a1ee878cec9e","owner":[],"postedDate":"February 23rd, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-04-25T10:08:56+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-23 06:58:55","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8833559","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8833559","identity":"rs-8833559","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}