Effect of treatment with GLP-1R agonists on the urinary peptidome of T2DM patients

Aim: Type II Diabetes mellitus (T2DM) accounts for approximately 90% of all DM cases in the world; and is diagnosed with increased levels of blood glucose or hyperglycemia. Glucagon-like peptide-1 receptor (GLP-1R) agonists have established an increased capability to target directly or indirectly six core defects associated with T2DM, while, the underlying molecular mechanisms of these pharmacological effects are not fully known. This exploratory study was conducted to analyze the effect of treatment with GLP-1R agonists on the urinary peptidome of T2DM patients. Material and Methods: Urine samples of thirty-two T2DM patients from the PROVALID study (A Prospective Cohort Study in Patients with T2DM for Validation of Biomarkers) were collected at pre- and post-treatment with GLP-1R agonist drugs, and analyzed by capillary electrophoresis coupled to mass spectrometry (CE-MS). Results: In total, 329 urinary peptides were identified, of which 70 were significantly affected by the GLP-1R agonist treatment; fragmenting from 26 different proteins. The downregulation of MMP proteases, based on the concordant downregulation of urinary collagen peptides with GLP-1R agonist treatment was highlighted in the study. Treatment also resulted in the downregulation of peptides from SERPINA1, APOC3, CD99, CPSF6, CRNN, SERPINA6, HBA2, MB, VGF, PIGR and TTR, many of which were previously found associated with increased kidney and/or vascular damage, indicating beneficial effect of GLP-1R agonists. Conclusions: The novel findings in this study, indicate potential molecular mechanisms of GLP-1R agonists in the context of management of T2DM and prevention or delaying the progression of its associated diseases. Keywords: T2DM, GLP-1R agonists, CE-MS, peptidomics, urine, peptide, collagen, COL1A1, COL3A1, MMP ### Competing Interest Statement The authors declare no conflict of interest. Author H.M. is the founder and co-owner of Mosaiques Diagnostics (Hannover, Germany). Authors E.M. and J.S. are employed by Mosaiques Diagnostics. ### Clinical Protocols ### Funding Statement This work was supported by the European Union Horizon 2020 research and innovation program by grant No 860329 (Marie-Curie ITN STRATEGY-CKD) and No 848011 (DC-ren). ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The PROVALID study protocol was approved in each participating country by the responsible local Institutional Review Board (IRB). Signing an informed consent was a prerequisite for study participation in all countries. Participating countries: Austria, Hungary, the Netherlands, Scotland, Poland. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors.