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Clinical Trial
. 2025 Jul;22(4):e00591.
doi: 10.1016/j.neurot.2025.e00591. Epub 2025 Apr 23.

Evaluation of exploratory fluid biomarkers from a phase 1 senolytic trial in mild Alzheimer's disease

Valentina R Garbarino  1 , Juan Pablo Palavicini  2 , Justin Melendez  3 , Nicolas R Barthelemy  3 , Yingxin He  3 , Tiffany F Kautz  4 , Marisa Lopez-Cruzan  5 , Julia J Mathews  6 , Peng Xu  7 , Bin Zhang  7 , Afaf Saliba  8 , Nagarjunachary Ragi  8 , Kumar Sharma  8 , Dallin Mason  9 , Samuel Johnson  9 , Suzanne Hendrix  9 , Suzanne Craft  10 , Ronald C Petersen  11 , Jair Machado Espindola-Netto  12 , Ailing Xue  12 , Tamara Tchkonia  12 , James L Kirkland  13 , Arash Salardini  14 , Nicolas Musi  15 , Randall J Bateman  3 , Mitzi M Gonzales  16 , Miranda E Orr  17
Affiliations
Clinical Trial

Evaluation of exploratory fluid biomarkers from a phase 1 senolytic trial in mild Alzheimer's disease

Valentina R Garbarino et al. Neurotherapeutics. 2025 Jul.

Abstract

Senescent cell accumulation contributes to the progression of age-related disorders including Alzheimer's disease (AD). Clinical trials focused on cellular senescence are in early stages and have yet to establish reliable outcome measures reflecting senescent cell burden or response to senolytics, therapeutics that clear senescent cells. Results from the first open-label trial of senolytics, dasatinib plus quercetin (D ​+ ​Q), in older adults (N ​= ​5) with early AD demonstrated central nervous system penetration of dasatinib and favorable safety and tolerability. Herein, we present exploratory analyses of senescence and AD-associated analytes in blood, cerebrospinal fluid (CSF) and urine from this study in effort to guide biomarker development for future senolytic trials. Immunoassays, mass spectrometry and transcriptomics were performed and changes in analyte levels were assessed from baseline to post-treatment using paired t-tests. Targeted cytokine and chemokine analyses revealed increases in plasma fractalkine and MMP-7 and CSF IL-6 from baseline to post-treatment. Mass spectrometry indicated stable levels of amyloid β and tau proteins in CSF, unchanged urinary metabolites, and modest treatment-associated lipid profile changes. Targeted transcriptomic analysis of peripheral blood mononuclear cells indicated downregulation of inflammatory genes including FOS, FOSB, IL1β, IL8, JUN, JUNB, PTGS2. The levels and treatment responses of the analytes identified here may help inform trial design and outcomes for senolytic studies. Independent validation will be necessary to develop standardized biomarker panels across senolytic trials for AD. ClinicalTrials.gov: NCT04063124.

Keywords: Alzheimer’s disease; Biofluids; Biomarkers; Clinical trial; Senolytics.

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Conflict of interest statement

Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ronald C. Petersen reports a relationship with University of Oxford, UpToDate, and Medscape that includes equity or stocks. Ronald C. Petersen reports personal stock in AbbVie. reports personal fees from Roche, Genetech, Eli Lilly, and Nestle, and no personal fees from Eisai, outside of the submitted work. Suzanne Craft reports a relationship with TD3 Therapeutics and Neurodegenerative Consortium that includes board membership. Suzanne Craft reports a relationship with vTv Therapeutics, Cylcerion, T3D Therapeutics, and Cognito Therapeutics, outside the submitted work that includes equity or stock. Randall J. Bateman reports a relationship with C2N Diagnostics and receives income from serving on the scientific advisory board as a co-founder. Randall J. Bateman has received research funding from Avid Radiopharmaceuticals, Janssen, Roche/Genentech, Eli Lilly, Eisai, Biogen, AbbVie, Bristol Myers Squibb, and Novartis. Mitzi M. Gonzales reports personal stock in Abbvie. James L. Kirkland and Tamara Tchkonia are co-investigators on a patent Treating Cognitive Decline and Other Neurodegenerative Conditions by Selectively Removing Senescent Cells from Neurological Tissue and a patent for Treating Cognitive Decline and Other Neurodegenerative Conditions by Selectively Removing Senescent Cells from Neurological Tissue that are held by Mayo Clinic with royalties paid to Mayo Clinic by Unity Biotechnologies. Dallin Mason, Samuel Johnson, and Suzanne Hendrix are employees of Pentara Corporation which provides consulting to over 30 pharmaceutical, biotech, non-profit, and academic groups doing clinical research in neurodegenerative disorders. Suzanne Hendrix is the founder, owner, and CEO of Pentara Corporation. Washington University has equity ownership interest in C2N Diagnostics and receives royalty income based on technology (Stable Isotope Labeling Kinetics, Blood Plasma Assay, and Methods of Diagnosing AD with Phosphorylation Changes) licensed by Washington University to C2N Diagnostics. Miranda Orr has patent Biosignature and Therapeutic Approach for Neuronal Senescence pending. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Inflammatory protein levels altered by dasatinib plus quercetin (D ​+ ​Q) treatment measured by Luminex® protein platform. a–d, Effect of dasatinib plus quercetin (D ​+ ​Q) on plasma and cerebrospinal fluid (CSF) inflammatory markers. Mean difference (95 ​% CI): a, plasma fractalkine, 629 ​mg/ml (549.90–705.60); b, plasma MMP-7, 226 ​mg/ml (0.198–452.90); c, CSF IL-6, 1.06 ​pg ​ml−1 (0.500–1.616). Baseline to post-treatment changes were assessed using two-sided paired sample t-tests, P ​< ​0.05, N ​= ​3–5, color coded by participant. Mean difference ​= ​post-treatment - baseline; 95 ​% CI, for the post versus baseline mean difference. No correction for multiple comparisons was made due to small sample size.
Fig. 2
Fig. 2
Correlation between cerebrospinal fluid (CSF) dasatinib (D) versus neurofilament light chain (NfL) levels. Post-treatment dasatinib (D; ng/ml) level correlation with baseline cerebrospinal fluid neurofilament light chain (NfL; pg/ml) derived from simple linear regression. R2 ​= ​0.7373; P ​= ​0.0624.
Fig. 3
Fig. 3
Effects of dasatinib plus quercetin (D ​+ ​Q) treatment on the circulating plasma lipidome normalized to total protein concentration. a–d, Effects of dasatinib plus quercetin (D ​+ ​Q) treatment on the circulating plasma lipidome normalized to total protein content. Plasma lipidome was assessed using multidimensional mass spectrometry-based shotgun lipidomics. a, MetaboAnalyst unsupervised PCA plot reducing all plasma lipid species data into three dimensions. Baseline and post-treatment groups are color-coded in gray and orange respectively, subjects are color coded to match color code assignments across all figures. b, All 11 lipid classes assessed in plasma samples. Paired samples are connected with a line. c, Volcano plot comparing all 194 plasma lipid species at baseline and post-treatment. d, Plot of the nine differentially abundant lipids (DALs) lipid species significantly decreased from baseline to post-treatment. Paired samples are connected with a line, each color represents a different subject (N ​= ​5). Only P ​< ​0.1 are shown.
Fig. 4
Fig. 4
Baseline and post-treatment significantly differentially expressedconservedtranscriptionalresponse toadversity (CTRA) gene counts in peripheral blood mononuclear cell samples measured with nanoString nCounter XT CodeSet custom CTRA gene expression panel. a–g, Effects of dasatinib plus quercetin (D ​+ ​Q) on the expression of Conserved Transcriptional Response to Adversity (CTRA) gene counts measured in peripheral blood mononuclear cell (PBMC) samples. Seven inflammatory genes were significantly decreased post-treatment. Log2 fold-change (B-statistic): a, FOSB, −2.176 counts (−0.963); b, PTGS2, -1.980 counts (−1.395); c, IL-8, -2.439 counts (−1.436) (d) FOS, −1.584 counts (−1.849); (e) IL-1B, −1.386 counts (−2.082), (f) JUNB, −0.859 counts (−3.605) (g) JUN, −0.969 counts (−3.808). Baseline to post-treatment changes were assessed using a moderated t-test by Robust Empirical Bayes analysis in limma, P ​< ​0.05, N ​= ​2–4, color coded by participant. Paired baseline and post-treatment measures existed for all but one of the participants (blue) for whom only a post-treatment PBMC sample was collected. No correction for multiple comparisons was made due to small sample size.
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