V. Suryadevara , R. von Kruechten , J.H. Tang , A.M. Dreisbach , Z. Shokri Varniab , S.B. Singh , A. Lubke , T. Liang , J. Wong , J. Wang , R. Duwa , J. Wang , M. Barbieri , F. Kogan , S.B. Goodman , L. Chou , D. Oji , J. Chan , T.J. Meade , H.E. Daldrup-Link
{"title":"CLINICAL TRANSLATION PIPELINE FOR DETECTING SENESCENCE IN OSTEOARTHRITIS USING THE Β-GALACTOSIDASE RESPONSIVE GD-CHELATE","authors":"V. Suryadevara , R. von Kruechten , J.H. Tang , A.M. Dreisbach , Z. Shokri Varniab , S.B. Singh , A. Lubke , T. Liang , J. Wong , J. Wang , R. Duwa , J. Wang , M. Barbieri , F. Kogan , S.B. Goodman , L. Chou , D. Oji , J. Chan , T.J. Meade , H.E. Daldrup-Link","doi":"10.1016/j.ostima.2025.100335","DOIUrl":null,"url":null,"abstract":"<div><h3>INTRODUCTION</h3><div>Cellular senescence is one of the key mechanisms implicated in the development and progression of OA. The identification of senescence-mediated molecular mechanisms in OA needs novel imaging tools to detect senescence and monitor the efficacy of new senolytic therapies. Progress in molecular imaging techniques has led to the creation of a novel β-gal responsive Gd-chelate for identifying senescence using MRI, the widely used imaging modality for OA.</div></div><div><h3>OBJECTIVE</h3><div>The hypothesis is that β-gal responsive Gd-chelate can detect senescence <em>in vitro, in vivo</em> and in human OA specimens.</div></div><div><h3>METHODS</h3><div>Senescence was induced in mesenchymal stem cells (MSCs) using 400nM doxorubicin over 5 days. Control and senescent cell suspensions incubated with 0.25 mM β-gal responsive Gd-chelate underwent MRI on a 3T MRI scanner (Bruker BioSpec, Billerica, MA). Further cartilage defects created in pig knees were implanted with control and senescent cells, followed by MRI after intraarticular injection of 2.5 mM β-gal responsive Gd-chelate. As a first step of clinical translation, human OA specimens were obtained from 30 patients undergoing hip/knee/ankle replacement. The fresh specimens were incubated with 2.5 mM of β-gal responsive Gd-chelate for an hour, before and after which MRI was performed using the following parameters: fat-saturated PD-weighted fast spin-echo sequence (TR=1500ms, matrix size=512 × 512pixels, slice thickness(=1mm, FOV=15cm, and NEX=2); SMART1 MAP sequence (TR = 40, 75, 150, 300, 500, 700, and 2,000 ms, matrix size=160 × 160 pixels SL=6mm, FOV=15 cm, and NEX=1) and T<sub>1</sub> weighted fast SE sequence (TR=500ms, matrix size=512 × 512 pixels, SL=1mm, FOV=15cm, and NEX=2). T1 maps were generated to calculate the T1 relaxation times.</div></div><div><h3>RESULTS</h3><div>Senescence was first confirmed with immunohistochemistry for senescence markers including p16, p21 and β-gal. <em>In vitro</em> studies indicated that senescent MSCs demonstrated a notable increase in MRI signal after being incubated with the β-gal responsive Gd-chelate probe, compared to control cells (Fig. 1A). <em>In vivo</em>, the probe was injected intraarticularly into pig knee joints, and a marked decrease in T1 relaxation times indicated the retention of the probe and it’s activation by senescent cells in cartilage defects (Fig. 1B). The Wilcoxon ranksum test was used to determine the significance between control and senescence group. In human OA specimens, areas with severe cartilage damage as graded by a radiologist using Outerbridge score demonstrated higher number of senescent cells seen on immunohistochemistry. MRI indicated that there is pronounced hyperintense signal in the T1-SE images upon incubation with the β-gal responsive Gd-chelate probe, compared to MRI of the specimens before incubation. This was further quantified on T1 maps and indicated a significant reduction in T<sub>1</sub> relaxation times, which also correlated with the Outerbridge score (Fig. 1C). The ordinal logistic regression indicated a significant negative correlation between T<sub>1</sub> relaxation times and Outerbridge score (p<0.0001).</div></div><div><h3>CONCLUSIONS</h3><div>This study demonstrates the clinical translation of the β-gal responsive Gd-chelate for detecting senescent cells <em>in vitro, in vivo</em> and in human OA specimens.</div></div>","PeriodicalId":74378,"journal":{"name":"Osteoarthritis imaging","volume":"5 ","pages":"Article 100335"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Osteoarthritis imaging","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2772654125000753","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
INTRODUCTION
Cellular senescence is one of the key mechanisms implicated in the development and progression of OA. The identification of senescence-mediated molecular mechanisms in OA needs novel imaging tools to detect senescence and monitor the efficacy of new senolytic therapies. Progress in molecular imaging techniques has led to the creation of a novel β-gal responsive Gd-chelate for identifying senescence using MRI, the widely used imaging modality for OA.
OBJECTIVE
The hypothesis is that β-gal responsive Gd-chelate can detect senescence in vitro, in vivo and in human OA specimens.
METHODS
Senescence was induced in mesenchymal stem cells (MSCs) using 400nM doxorubicin over 5 days. Control and senescent cell suspensions incubated with 0.25 mM β-gal responsive Gd-chelate underwent MRI on a 3T MRI scanner (Bruker BioSpec, Billerica, MA). Further cartilage defects created in pig knees were implanted with control and senescent cells, followed by MRI after intraarticular injection of 2.5 mM β-gal responsive Gd-chelate. As a first step of clinical translation, human OA specimens were obtained from 30 patients undergoing hip/knee/ankle replacement. The fresh specimens were incubated with 2.5 mM of β-gal responsive Gd-chelate for an hour, before and after which MRI was performed using the following parameters: fat-saturated PD-weighted fast spin-echo sequence (TR=1500ms, matrix size=512 × 512pixels, slice thickness(=1mm, FOV=15cm, and NEX=2); SMART1 MAP sequence (TR = 40, 75, 150, 300, 500, 700, and 2,000 ms, matrix size=160 × 160 pixels SL=6mm, FOV=15 cm, and NEX=1) and T1 weighted fast SE sequence (TR=500ms, matrix size=512 × 512 pixels, SL=1mm, FOV=15cm, and NEX=2). T1 maps were generated to calculate the T1 relaxation times.
RESULTS
Senescence was first confirmed with immunohistochemistry for senescence markers including p16, p21 and β-gal. In vitro studies indicated that senescent MSCs demonstrated a notable increase in MRI signal after being incubated with the β-gal responsive Gd-chelate probe, compared to control cells (Fig. 1A). In vivo, the probe was injected intraarticularly into pig knee joints, and a marked decrease in T1 relaxation times indicated the retention of the probe and it’s activation by senescent cells in cartilage defects (Fig. 1B). The Wilcoxon ranksum test was used to determine the significance between control and senescence group. In human OA specimens, areas with severe cartilage damage as graded by a radiologist using Outerbridge score demonstrated higher number of senescent cells seen on immunohistochemistry. MRI indicated that there is pronounced hyperintense signal in the T1-SE images upon incubation with the β-gal responsive Gd-chelate probe, compared to MRI of the specimens before incubation. This was further quantified on T1 maps and indicated a significant reduction in T1 relaxation times, which also correlated with the Outerbridge score (Fig. 1C). The ordinal logistic regression indicated a significant negative correlation between T1 relaxation times and Outerbridge score (p<0.0001).
CONCLUSIONS
This study demonstrates the clinical translation of the β-gal responsive Gd-chelate for detecting senescent cells in vitro, in vivo and in human OA specimens.