Extracellular Vesicles in regenerative medicine

Benedetta Bussolati - PI

Benedetta Bussolati - Full Professor of Laboratory Medicine, Department of Molecular Biotechnology and Health Sciences, University of Torino, Italy

Main group members

Transplant Biology Group

Alessia Brossa RTDA Roberta Verta-PhD
Adele Tanzi PhD
Tunahan Ergünay PhD
Alessia Dalmasso PhD
Marta Fornaro PhD
Michela Arena PhD

Research activity

Our group has a strong background on renal pathophysiology, on angiogenesis and on the mechanisms of renal damage and progression. In addition, we have extensive experience in studies of stem cell biology and regenerative medicine that include characterization of various stem cell types and derived bioproducts and their potential use for tissue regeneration. In particular, we are investigating investigate the role of extracellular vesicles in regenerative medicine as delivery systems of therapeutic cargo, such as RNA species and as diagnostic tool. A platform for single-EV characterization with dedicated instruments is presentin the lab. Moreover, we are currently studying other cell derived bioproducts, such as mitochondria, for regenerative medicine, with a particular focus on graft protection during reperfusion.

EVs as diagnostic tools

We are interested in the characterization of urinary EVs in order to identify disease biomarkers, as well as markers of renal regeneration reserve. We characterized EVs in amniotic fluid, which is mainly derived from fetal urine, and identified their alteration in pre-eclamptic patients (Fig.2). We also extensively characterized urinary EVs These vesicles are of interest as mediators of glomerular-tubular and intersegment tubular crosstalk and are involved in the amplification of kidney damage and inflammation. The molecular profile of extracellular vesicles reflects the type and pathophysiological status of the originating cell so could potentially be exploited for diagnostic and prognostic purposes. In recent studies, we showed that extracellular vesicles present in urine may represent useful markers of renal regenerative ability of interest to assess the risk of chronic kidney disease progression in transplanted patients (Fig.3).

EVs as therapeutic tools

Extracellular vesicles from different stem cell types might be used for regeneration of renal tissue after injury. We showed that urinary EVs can contain Klotho, a single-pass transmembrane hormone identified as an antiaging factor, expressed in the kidney where it exherts a regenerative effect (Fig.5). Our results demonstrated the novel potential use of urinary EVs with factor Klotho to contrast the acute kidney injury Extracellular vesicles, especially from autologous sources, are also attractive candidates for drug delivery and various engineering strategies are being investigated to modify their cargo and increase their efficacy (Fig. 6). Finally, we recently demonstrated the regenerative effect of mitochondrial transplant in a model of ex vivo kidney reperfusion, resembling cardiac death donation.

Figure 2

Illustration of the multiple possible sources of term amniotic fluid -de - rived EVs and the main changes in preeclampsia- de - rived -EVs. Their antiangiogenic properties are supported by the specific upregulation of CD105 surface. (Gebara et al. JEV 2022)

Figure 5

Main functions of the Klotho protein

FUTURE RESEARCH PLANS

We aim to extend our results on urinary EVs and to identify an early signature of renal damage progression, to be transplanted into the clinic. We also aim to exploit EVs and mitochondria for organ regeneration, with a main focus on graf function.

FUNDING ID (PAST 5 YEARS)

2023-2026 PNRR National Center for the development of RNA-based therapies. Spoke 8.1 - Translational development of smart delivery platforms. PI of the 8.1.4 – Biological vesicles and platforms
2020-2025. NIH R01 Grant No. R01DK121037 Extracellular vesicles derived from amniotic fluid stem cells normalize glomerular function during progressive kidney disease, Co-PI.
2018-2023 European H2020 project RenalToolBox H2020-MSCA-ITN-2018
2016-2022 Unicyte EG AV Pre-clinical development of stem cell-derived EVs for treatment of Renal Carcinomas
2017-2021 European H2020 project iPLACENTA: H2020-MSCA-ITN-2017
2015-2020 Grant of the Italian Association for Cancer Research (AIRC), IG2015 169173

SELECTED PUBLICATIONS

Rossi A, Asthana A, Riganti C, Sedrakyan S, Byers LN, Robertson J, Senger RS, Montali F, Grange C, Dalmasso A, Porporato PE, Palles C, Thornton ME, Da Sacco S, Perin L, Ahn B, McCully J, Orlando G, Bussolati B. Mitochondria Transplantation Mitigates Damage in an In Vitro Model of Renal Tubular Injury and in an Ex Vivo Model of DCD Renal Transplantation. Ann Surg. 2023 Jul 14. doi: 10.1097/SLA.0000000000006005.
Burrello J, et al. Identification of a serum and urine ex - tracellular vesicle signature predicting renal outcome after kidney transplant. Nephrol Dial Transplant. 2023 28;38(3):764-777. doi: 10.1093/ndt/gfac259.
Grange C, Bussolati B. Extracellular vesicles in kidney disease. Nat Rev Nephrol. 2022;18:499-513. 10.1038/ s41581-022-00586-9
Gebara N, et al. Single extracellular vesicle analysis in human amniotic fluid shows evidence of phenotype alterations in preeclampsia. J Extracell Vesicles. 2022;11(5):e12217. doi: 10.1002/jev2.12217.
Börger V, et al. International Society for Extracellular Vesicles and International Society for Cell and Gene Therapy statement on extracellular vesicles from mesenchymal stromal cells and other cells: considerations for potential therapeutic agents to suppress coronavirus disease-19. Cytotherapy. 2020; 22(9):482-485. doi: 10.1016/j.jcyt.2020.05.002.
Bellucci L, et al. Mesenchymal Stromal Cell-Derived Extracellular Vesicles Pass through the Filtration Barrier and Protect Podocytes in a 3D Glomerular Model under Continuous Perfusion. Tissue Eng Regen Med. 2021;18(4):549-560. doi: 10.1007/s13770-021-00374-9.
Erdbrügger U, et al. Urinary extracellular vesicles: A position paper by the Urine Task Force of the Interna - tional Society for Extracellular Vesicles. J Extracell Vesi - cles. 2021;10(7):e12093. doi: 10.1002/jev2.12093.
Iampietro C, et al. Bussolati B. Molecular and functional characterization of urine-derived podocytes from patients with Alport syndrome. J Pathol. 2020; 252(1):88- 100. doi: 10.1002/path.5496.
Grange C, et al. Urinary Extracellular Vesicles Carrying Klotho Improve the Recovery of Renal Function in an Acute Tubular Injury Model. Mol Ther. 2020; 28(2):490- 502. doi: 10.1016/j.ymthe.2019.11.013
Lopatina T, et al. Extracellular vesicles from human liver stem cells inhibit tumor angiogenesis. Int J Cancer.2019;144(2):322-333.5. 6. doi: 10.1002/ijc.31796

Ultimo aggiornamento: 05/11/2024