Publications by authors named "Federica Casiraghi"

49 Publications

Lysophosphatidic Acid: Promoter of Cancer Progression and of Tumor Microenvironment Development. A Promising Target for Anticancer Therapies?

Cells 2021 Jun 4;10(6). Epub 2021 Jun 4.

Istituto di Ricerche Farmacologiche Mario Negri IRCCS, 24126 Bergamo, Italy.

Increased expression of the enzyme autotaxin (ATX) and the consequently increased levels of its product, lysophosphatidic acid (LPA), have been reported in several primary tumors. The role of LPA as a direct modulator of tumor cell functions-motility, invasion and migration capabilities as well as resistance to apoptotic death-has been recognized by numerous studies over the last two decades. Notably, evidence has recently been accumulating that shows that LPA also contributes to the development of the tumor microenvironment (TME). Indeed, LPA plays a crucial role in inducing angiogenesis and lymphangiogenesis, triggering cellular glycolytic shift and stimulating intratumoral fibrosis. In addition, LPA helps tumoral cells to escape immune surveillance. Treatments that counter the TME components, in order to deprive cancer cells of their crucial support, have been emerging among the promising new anticancer therapies. This review aims to summarize the latest knowledge on how LPA influences both tumor cell functions and the TME by regulating the activity of its different elements, highlighting why and how LPA is worth considering as a molecular target for new anticancer therapies.
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http://dx.doi.org/10.3390/cells10061390DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8229068PMC
June 2021

Chronic lung allograft pathology lesions in two rat strain combinations.

J Thorac Dis 2021 May;13(5):2833-2843

Department of Cardiac, Thoracic, Vascular Sciences and Public Health, University of Padova Medical School, Padova, Italy.

Background: Chronic lung allograft dysfunction remains an obstacle to long-term survival after lung transplantation. Two phenotypes have been described: obliterative bronchiolitis and restrictive allograft syndrome. Preclinical models are essential to analyze chronic lung allograft dysfunction pathophysiology.

Methods: Orthotopic lung transplants from 38 Lewis into Fischer 344 (Lew→F344) and 67 Brown-Norway into Lewis (BN→Lew) rats were performed in our center in the last decade. We carefully reviewed and quantified all grafts with chronic rejection (40 cases) (18 Lew→F344, 22 BN→Lew) with the aim to investigate if histological changes of chronic lung allograft dysfunction could be also detected in rat grafts.

Results: All animals showed human reminiscent histological lesions. Early chronic rejection lesions were detected in BN→Lew. End-stage chronic rejection with features of obliterative bronchiolitis was observed in 33% of Lew→F344; end-stage with restrictive allograft syndrome chronic rejection in 67% and 80% of Lew→F344 and BN→Lew, respectively. BN→Lew showed higher grades of endotheliitis, vascular fibrosis, and lower grades of lymphoid aggregates than Lew→F344 (P=0.007, P=0.043, P=0.004, respectively).

Conclusions: Chronic rejection lesions in rat lung allografts mimic those in humans. The frequent occurrence of restrictive allograft syndrome-like lesions in BN→Lew may be related to a higher degree of mismatch in this strain combination. These animal models could allow future mechanistic studies to better understand chronic lung allograft dysfunction pathogenesis.
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http://dx.doi.org/10.21037/jtd-20-3415DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8182524PMC
May 2021

Amnion epithelial cells are an effective source of factor H and prevent kidney complement deposition in factor H-deficient mice.

Stem Cell Res Ther 2021 06 10;12(1):332. Epub 2021 Jun 10.

Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Via GB Camozzi 3, 24020, Ranica, Bergamo, Italy.

Complement factor H (FH) is the main plasma regulator of the alternative pathway of complement. Genetic and acquired abnormalities in FH cause uncontrolled complement activation amplifying, with the consequent accumulation of complement components on the renal glomeruli. This leads to conditions such as C3 glomerulopathy (C3G) and atypical hemolytic uremic syndrome (aHUS). There is no effective therapy for these diseases. Half of the patients progress to end-stage renal disease and the condition recurs frequently in transplanted kidneys. Combined liver/kidney transplantation is a valid option for these patients, but the risks of the procedure and donor organ shortages hamper its clinical application. Therefore, there is an urgent need for alternative strategies for providing a normal FH supply. Human amnion epithelial cells (hAEC) have stem cell characteristics, including the capability to differentiate into hepatocyte-like cells in vivo.Here, we administered hAEC into the livers of newborn Cfh mice, which spontaneously developed glomerular complement deposition and renal lesions resembling human C3G. hAEC engrafted at low levels in the livers of Cfh mice and produced sufficient human FH to prevent complement activation and glomerular C3 and C9 deposition. However, long-term engraftment was not achieved, and eventually hAEC elicited a humoral immune response in immunocompetent Cfh mice.hAEC cell therapy could be a valuable therapeutic option for patients undergoing kidney transplantation in whom post-transplant immunosuppression may protect allogeneic hAEC from rejection, while allogeneic cells provide normal FH to prevent disease recurrence.
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http://dx.doi.org/10.1186/s13287-021-02386-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8194190PMC
June 2021

Mesenchymal Stromal Cell Therapy in Solid Organ Transplantation.

Front Immunol 2020 10;11:618243. Epub 2021 Feb 10.

Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Aldo & Cele Daccò Clinical Research Center for Rare Diseases, Bergamo, Italy.

Transplantation is the gold-standard treatment for the failure of several solid organs, including the kidneys, liver, heart, lung and small bowel. The use of tailored immunosuppressive agents has improved graft and patient survival remarkably in early post-transplant stages, but long-term outcomes are frequently unsatisfactory due to the development of chronic graft rejection, which ultimately leads to transplant failure. Moreover, prolonged immunosuppression entails severe side effects that severely impact patient survival and quality of life. The achievement of tolerance, i.e., stable graft function without the need for immunosuppression, is considered the Holy Grail of the field of solid organ transplantation. However, spontaneous tolerance in solid allograft recipients is a rare and unpredictable event. Several strategies that include peri-transplant administration of non-hematopoietic immunomodulatory cells can safely and effectively induce tolerance in pre-clinical models of solid organ transplantation. Mesenchymal stromal cells (MSC), non-hematopoietic cells that can be obtained from several adult and fetal tissues, are among the most promising candidates. In this review, we will focus on current pre-clinical evidence of the immunomodulatory effect of MSC in solid organ transplantation, and discuss the available evidence of their safety and efficacy in clinical trials.
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http://dx.doi.org/10.3389/fimmu.2020.618243DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7902912PMC
June 2021

Third-party bone marrow-derived mesenchymal stromal cell infusion before liver transplantation: A randomized controlled trial.

Am J Transplant 2020 Dec 28. Epub 2020 Dec 28.

Aldo & Cele Daccò Clinical Research Center for Rare Diseases, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Bergamo, Italy.

Mesenchymal stromal cells (MSC) have emerged as a promising therapy to minimize the immunosuppressive regimen or induce tolerance in solid organ transplantation. In this randomized open-label phase Ib/IIa clinical trial, 20 liver transplant patients were randomly allocated (1:1) to receive a single pretransplant intravenous infusion of third-party bone marrow-derived MSC or standard of care alone. The primary endpoint was the safety profile of MSC administration during the 1-year follow-up. In all, 19 patients completed the study, and none of those who received MSC experienced infusion-related complications. The incidence of serious and non-serious adverse events was similar in the two groups. Circulating Treg/memory Treg and tolerant NK subset of CD56 NK cells increased slightly over baseline, albeit not to a statistically significant extent, in MSC-treated patients but not in the control group. Graft function and survival, as well as histologic parameters and intragraft expression of tolerance-associated transcripts in 1-year protocol biopsies were similar in the two groups. In conclusion, pretransplant MSC infusion in liver transplant recipients was safe and induced mild positive changes in immunoregulatory T and NK cells in the peripheral blood. This study opens the way for a trial on possible tolerogenic efficacy of MSC in liver transplantation. ClinicalTrials.gov identifier: NCT02260375.
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http://dx.doi.org/10.1111/ajt.16468DOI Listing
December 2020

Cellular therapies in organ transplantation.

Transpl Int 2021 02 15;34(2):233-244. Epub 2021 Jan 15.

Nephrology and Transplantation, Department of Internal Medicine, Erasmus University Medical Center, Erasmus Medical Center, Rotterdam, The Netherlands.

Cellular therapy is a promising tool for improving the outcome of organ transplantation. Various cell types with different immunoregulatory and regenerative properties may find application for specific transplant rejection or injury-related indications. The current era is crucial for the development of cellular therapies. Preclinical models have demonstrated the feasibility of efficacious cell therapy in transplantation, early clinical trials have shown safety of several of these therapies, and the first steps towards efficacy studies in humans have been made. In this review, we address the current state of the art of cellular therapies in clinical transplantation and discuss monitoring tools and endpoints for these studies.
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http://dx.doi.org/10.1111/tri.13789DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7898347PMC
February 2021

Novel alternative ribonucleotide excision repair pathways in human cells by DDX3X and specialized DNA polymerases.

Nucleic Acids Res 2020 11;48(20):11551-11565

Institute of Molecular Genetics IGM-CNR 'Luigi Luca Cavalli-Sforza', via Abbiategrasso 207, I-27100 Pavia, Italy.

Removal of ribonucleotides (rNMPs) incorporated into the genome by the ribonucleotide excision repair (RER) is essential to avoid genetic instability. In eukaryotes, the RNaseH2 is the only known enzyme able to incise 5' of the rNMP, starting the RER process, which is subsequently carried out by replicative DNA polymerases (Pols) δ or ϵ, together with Flap endonuclease 1 (Fen-1) and DNA ligase 1. Here, we show that the DEAD-box RNA helicase DDX3X has RNaseH2-like activity and can support fully reconstituted in vitro RER reactions, not only with Pol δ but also with the repair Pols β and λ. Silencing of DDX3X causes accumulation of rNMPs in the cellular genome. These results support the existence of alternative RER pathways conferring high flexibility to human cells in responding to the threat posed by rNMPs incorporation.
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http://dx.doi.org/10.1093/nar/gkaa948DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672437PMC
November 2020

Immunity, endothelial injury and complement-induced coagulopathy in COVID-19.

Nat Rev Nephrol 2021 01 19;17(1):46-64. Epub 2020 Oct 19.

Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Bergamo, Italy.

In December 2019, a novel coronavirus was isolated from the respiratory epithelium of patients with unexplained pneumonia in Wuhan, China. This pathogen, named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causes a pathogenic condition that has been termed coronavirus disease 2019 (COVID-19) and has reached pandemic proportions. As of 17 September 2020, more than 30 million confirmed SARS-CoV-2 infections have been reported in 204 different countries, claiming more than 1 million lives worldwide. Accumulating evidence suggests that SARS-CoV-2 infection can lead to a variety of clinical conditions, ranging from asymptomatic to life-threatening cases. In the early stages of the disease, most patients experience mild clinical symptoms, including a high fever and dry cough. However, 20% of patients rapidly progress to severe illness characterized by atypical interstitial bilateral pneumonia, acute respiratory distress syndrome and multiorgan dysfunction. Almost 10% of these critically ill patients subsequently die. Insights into the pathogenic mechanisms underlying SARS-CoV-2 infection and COVID-19 progression are emerging and highlight the critical role of the immunological hyper-response - characterized by widespread endothelial damage, complement-induced blood clotting and systemic microangiopathy - in disease exacerbation. These insights may aid the identification of new or existing therapeutic interventions to limit the progression of early disease and treat severe cases.
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http://dx.doi.org/10.1038/s41581-020-00357-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7570423PMC
January 2021

Unique Domain for a Unique Target: Selective Inhibitors of Host Cell DDX3X to Fight Emerging Viruses.

J Med Chem 2020 09 18;63(17):9876-9887. Epub 2020 Aug 18.

Istituto di Genetica Molecolare IGM-CNR "Luigi Luca Cavalli-Sforza", Via Abbiategrasso 207, I-27100 Pavia, Italy.

Emerging viruses like dengue, West Nile, chikungunya, and Zika can cause widespread viral epidemics. Developing novel drugs or vaccines against specific targets for each virus is a difficult task. As obligate parasites, all viruses exploit common cellular pathways, providing the possibility to develop broad-spectrum antiviral agents targeting host factors. The human DEAD-box RNA helicase DDX3X is an essential cofactor for viral replication but dispensable for cell viability. Herein, we exploited the presence of a unique structural motif of DDX3X not shared by other cellular enzymes to develop a theoretical model to aid in the design of a novel class of highly selective inhibitors acting against such specific targets, thus limiting off-targeting effects. High-throughput virtual screening led us to identify hit compound , endowed with promising antienzymatic activity. To improve its aqueous solubility, and its two enantiomers were synthesized and converted into their corresponding acetate salts (compounds , , and ). mutagenesis and biochemical and cellular assays further confirmed that the developed molecules were selective for DDX3X and were able to suppress replication of West Nile and dengue viruses in infected cells in the micromolar range while showing no toxicity for uninfected cells. These results provide proof of principle for a novel strategy in developing highly selective and broad-spectrum antiviral molecules active against emerging and dangerous viral pathogens. This study paves the way for the development of larger focused libraries targeting such domain to expand SAR studies and fully characterize their mode of interaction.
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http://dx.doi.org/10.1021/acs.jmedchem.0c01039DOI Listing
September 2020

The emergence of regenerative medicine in organ transplantation: 1st European Cell Therapy and Organ Regeneration Section meeting.

Transpl Int 2020 08 28;33(8):833-840. Epub 2020 Apr 28.

Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.

Regenerative medicine is emerging as a novel field in organ transplantation. In September 2019, the European Cell Therapy and Organ Regeneration Section (ECTORS) of the European Society for Organ Transplantation (ESOT) held its first meeting to discuss the state-of-the-art of regenerative medicine in organ transplantation. The present article highlights the key areas of interest and major advances in this multidisciplinary field in organ regeneration and discusses its implications for the future of organ transplantation.
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http://dx.doi.org/10.1111/tri.13608DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7497223PMC
August 2020

Transplantation-Induced Ischemia-Reperfusion Injury Modulates Antigen Presentation by Donor Renal CD11cF4/80 Macrophages through IL-1R8 Regulation.

J Am Soc Nephrol 2020 03 27;31(3):517-531. Epub 2020 Jan 27.

Department of Molecular Medicine, Istituto di Ricerche Farmacologiche Mario Negri Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Bergamo, Italy; and

Background: In donor kidneys subjected to ischemia-reperfusion injury during kidney transplant, phagocytes coexpressing the F4/80 and CD11c molecules mediate proinflammatory responses and trigger adaptive immunity in transplantation through antigen presentation. After injury, however, resident renal macrophages coexpressing these surface markers acquire a proreparative phenotype, which is pivotal in controlling inflammation and fibrosis. No data are currently available regarding the effects of transplant-induced ischemia-reperfusion injury on the ability of donor-derived resident renal macrophages to act as professional antigen-presenting cells.

Methods: We evaluated the phenotype and function of intragraft CD11cF4/80 renal macrophages after cold ischemia. We also assessed the modifications of donor renal macrophages after reversible ischemia-reperfusion injury in a mouse model of congeneic renal transplantation. To investigate the role played by IL-1R8, we conducted and studies comparing cells and grafts from wild-type and IL-R8-deficient donors.

Results: Cold ischemia and reversible ischemia-reperfusion injury dampened antigen presentation by renal macrophages, skewed their polarization toward the M phenotype, and increased surface expression of IL-1R8, diminishing activation mediated by toll-like receptor 4. Ischemic IL-1R8-deficient donor renal macrophages acquired an M phenotype, effectively induced IFN and IL-17 responses, and failed to orchestrate tissue repair, resulting in severe graft fibrosis and aberrant humoral immune responses.

Conclusions: IL-1R8 is a key regulator of donor renal macrophage functions after ischemia-reperfusion injury, crucial to guiding the phenotype and antigen-presenting role of these cells. It may therefore represent an intriguing pathway to explore with respect to modulating responses against autoantigens and alloantigens after kidney transplant.
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http://dx.doi.org/10.1681/ASN.2019080778DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7062225PMC
March 2020

Kidney transplant tolerance associated with remote autologous mesenchymal stromal cell administration.

Stem Cells Transl Med 2020 04 24;9(4):427-432. Epub 2019 Dec 24.

Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Bergamo, Italy.

Here we report the case of successful immune tolerance induction in a living-donor kidney transplant recipient remotely treated with autologous bone marrow-derived mesenchymal stromal cells (MSC). This case report, which to the best of our knowledge is the first in the world in this setting, provides evidence that the modulation of the host immune system with MSC can enable the safe withdrawal of maintenance immunosuppressive drugs while preserving optimal long-term kidney allograft function.
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http://dx.doi.org/10.1002/sctm.19-0185DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7103624PMC
April 2020

Update on mesenchymal stromal cell studies in organ transplant recipients.

Curr Opin Organ Transplant 2020 02;25(1):27-34

Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Bergamo.

Purpose Of Review: Mesenchymal stromal cells (MSC) have been proposed as a novel cell therapy for immune-mediated diseases, including solid organ transplantation. Here, we provide an overview of recent preclinical and clinical studies in solid organ transplantation using MSC immunomodulatory therapy.

Recent Findings: MSC have been tested successfully in models of corneal and lung transplantation, suggesting that either the programmed cell death protein 1/programmed death ligand 1 pathway or the generation of intermediary immune-regulatory monocyte-macrophage population are the main mechanisms of the protolerogenic effect of MSC. In clinical transplantation, allogeneic MSC from bone marrow or umbilical cord have been evaluated in kidney and lung transplantation with an excellent safety profile. Recent data from kidney transplant patients given autologous bone marrow-MSC enrolled in our phase 1 study demonstrated a good long term safety profile. Extensive immunomonitoring of this initial cohort provided evidence of the development of a protolerogenic environment in some MSC-treated patients. One of these patients has been weaned off immunosuppression successfully.

Summary: The available clinical studies in kidney, liver and lung transplantation indicate that autologous and allogeneic MSC therapy from different sources are safe. Now it's time to focus on well-designed efficacy clinical trials, possibly including extensive immunomonitoring.
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http://dx.doi.org/10.1097/MOT.0000000000000716DOI Listing
February 2020

Vein Suturing Results in Worse Lung Graft Outcomes Compared to the Cuff Method.

Eur Surg Res 2019 3;60(3-4):106-116. Epub 2019 Sep 3.

Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Bergamo, Italy,

Background: The rat orthotopic lung transplant model is not widely used yet because of the complexity of the procedure, in particular, venous anastomosis. Here, we performed a rat orthotopic lung transplantation using either the suture (ST) or cuff (CT) method for vein anastomosis.

Objectives: To compare the vein ST and CT techniques in terms of operative time, success, recipient survival, and early histological outcomes was the objective of this study.

Methods: A total of 24 left lung transplants in rats were performed. Twelve syngeneic (Lewis to Lewis) and 12 allogeneic (Brown-Norway to Lewis) lung transplants were performed using either the vein ST or the CT procedure. Arterial and bronchial anastomoses were performed with the CT technique. Graft histological damage was evaluated 3-7 days post-transplant in all rat lungs.

Results: The surgical success rate was 75% in both the ST and CT groups. Failures related mainly to vein bleeding (n = 2 in the ST group) and thrombosis (n = 1 in the ST group; n = 2 in the CT group). Total ischemia time was longer in the ST group (122 ± 25 min in ST group vs. 83 ± 10 min in CT group, mean ± SD), due to prolonged warm ischemia time (60 ± 12 min in the ST group vs. 21 ± 5 min in the CT group, mean ± SD), reflecting the time required to complete the vein ST procedure. The prolonged warm ischemia time resulted in significantly higher vascular inflammation in syngeneic grafts (2.3 ± 1.2 ST group vs. 0 in the CT group, mean ± SD) and in increased severity of ischemia/reperfusion injury and acute graft rejection (3.6 ± 0.4 in the ST group vs. 2.6 ± 0.4 in the CT group, mean ± SD) in allogeneic lung transplants.

Conclusions: The vein ST technique is a more time-consuming procedure than the CT method and the prolonged anastomosis time has a deleterious impact on transplant outcomes. These findings suggest that warm ischemia time - one of the modifiable transplant factors - should be considered a major risk factor in lung transplantation, particularly in the setting of donation after cardiac death.
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http://dx.doi.org/10.1159/000501805DOI Listing
March 2020

Mesenchymal Stromal Cells for Transplant Tolerance.

Front Immunol 2019 4;10:1287. Epub 2019 Jun 4.

Department of Molecular Medicine, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Bergamo, Italy.

In solid organ transplantation lifelong immunosuppression exposes transplant recipients to life-threatening complications, such as infections and malignancies, and to severe side effects. Cellular therapy with mesenchymal stromal cells (MSC) has recently emerged as a promising strategy to regulate anti-donor immune responses, allowing immunosuppressive drug minimization and tolerance induction. In this review we summarize preclinical data on MSC in solid organ transplant models, focusing on potential mechanisms of action of MSC, including down-regulation of effector T-cell response and activation of regulatory pathways. We will also provide an overview of available data on safety and feasibility of MSC therapy in solid organ transplant patients, highlighting the issues that still need to be addressed before establishing MSC as a safe and effective tolerogenic cell therapy in transplantation.
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http://dx.doi.org/10.3389/fimmu.2019.01287DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6559333PMC
October 2020

Effect of Timing and Complement Receptor Antagonism on Intragraft Recruitment and Protolerogenic Effects of Mesenchymal Stromal Cells in Murine Kidney Transplantation.

Transplantation 2019 06;103(6):1121-1130

IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Bergamo, Italy.

Background: Mesenchymal stromal cells (MSCs) have protolerogenic effects in renal transplantation, but they induce long-term regulatory T cells (Treg)-dependent graft acceptance only when infused before transplantation. When given posttransplant, MSCs home to the graft where they promote engraftment syndrome and do not induce Treg. Unfortunately, pretransplant MSC administration is unfeasible in deceased-donor kidney transplantation.

Methods: To make MSCs a therapeutic option also for deceased organ recipients, we tested whether MSC infusion at the time of transplant (day 0) or posttransplant (day 2) together with inhibition of complement receptors prevents engraftment syndrome and allows their homing to secondary lymphoid organs for promoting tolerance. We analyzed intragraft and splenic MSC localization, graft survival, and alloimmune response in mice recipients of kidney allografts and syngeneic MSCs given on day 0 or on posttransplant day 2. C3a receptor (C3aR) or C5a receptor (C5aR) antagonists were administered to mice in combination with the cells or were used together to treat MSCs before infusion.

Results: Syngeneic MSCs given at day 0 homed to the spleen increased Treg numbers and induced long-term graft acceptance. Posttransplant MSC infusion, combined with a short course of C3aR or C5aR antagonist or administration of MSCs pretreated with C3aR and C5aR antagonists, prevented intragraft recruitment of MSCs and graft inflammation, inhibited antidonor T-cell reactivity, but failed to induce Treg, resulting in mild prolongation of graft survival.

Conclusions: These data support testing the safety/efficacy profile of administering MSCs on the day of transplant in deceased-donor transplant recipients and indicate that complement is crucial for MSC recruitment into the kidney allograft.
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http://dx.doi.org/10.1097/TP.0000000000002611DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6934941PMC
June 2019

Mesenchymal stromal cells in kidney transplantation.

Curr Opin Nephrol Hypertens 2019 01;28(1):40-46

Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Bergamo, Italy.

Purpose Of Review: Mesenchymal stromal cells (MSC) have emerged as one of the most promising candidates for immunomodulatory cell therapy in kidney transplantation. Here we describe novel insights into the MSC mechanism of action and provide an overview of initial safety and feasibility studies with MSC in kidney transplantation.

Recent Findings: Clinical studies of MSC-based cell therapy in kidney transplant recipients demonstrated the safety and feasibility of cell therapy and provide the first encouraging evidence of the efficacy of MSC in enabling the minimization of immunosuppressive drugs. In our initial experience with MSC-based therapy in kidney transplant recipients we carried out extensive clinical and immunological monitoring of MSC-treated patients and found possible biomarkers of MSC immunomodulation in some of them. Based on these biomarkers we identified a patient in whom complete discontinuation of immunosuppression has been achieved safely and successfully.

Summary: Many issues should be addressed before MSC-based therapy becomes a standard treatment protocol for kidney transplantation. A better understanding of the MSC mechanism of action and the identification of biomarkers of response to therapy will inform the rational design of the most effective clinical protocol and the selection of patients amenable to safe immunosuppressive drug withdrawal.
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http://dx.doi.org/10.1097/MNH.0000000000000461DOI Listing
January 2019

Long-Term Clinical and Immunological Profile of Kidney Transplant Patients Given Mesenchymal Stromal Cell Immunotherapy.

Front Immunol 2018 14;9:1359. Epub 2018 Jun 14.

IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Bergamo, Italy.

We report here the long-term clinical and immunological results of four living-donor kidney transplant patients given autologous bone marrow-derived mesenchymal stromal cells (MSCs) as part of a phase 1 study focused on the safety and feasibility of this cell therapy. According to study protocols implemented over time, based on initial early safety findings, the patients were given MSC at day 7 posttransplant ( = 2) or at day -1 pretransplant ( = 2) and received induction therapy with basiliximab and low-dose rabbit anti-thymocyte globulin (RATG) or RATG alone, and were maintained on low-dose ciclosporin (CsA)/mycophenolate mofetil (MMF). All MSC-treated patients had stable graft function during the 5- to 7-year follow-up, without increased susceptibility to infections or neoplasm. In three MSC recipients, but not historical control patients, circulating memory CD8 T cell percentages remained lower than basal, coupled with persistent reduction of donor-specific cytotoxicity. Two patients showed a long-lasting increase in the regulatory T cell/memory CD8 T cell ratio, paralleled by high circulating levels of naïve and transitional B cells. In one of these two patients, CsA was successfully discontinued, and currently the low-dose MMF monotherapy is on the tapering phase. The study shows that MSC therapy is safe in the long term and could promote a pro-tolerogenic environment in selected patients. Extensive immunomonitoring of MSC-treated kidney transplant recipients could help selection of patients for safe withdrawal of maintenance immunosuppressive drugs (NCT00752479 and NCT02012153).
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http://dx.doi.org/10.3389/fimmu.2018.01359DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6014158PMC
June 2018

Mesenchymal stromal cells for tolerance induction in organ transplantation.

Hum Immunol 2018 May 27;79(5):304-313. Epub 2017 Dec 27.

IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Bergamo, Italy; Unit of Nephrology and Dialysis, Azienda Socio Sanitaria Territoriale (ASST), Papa Giovanni XXIII, Bergamo, Italy; L. Sacco Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy. Electronic address:

The primary challenge in organ transplantation continues to be the need to suppress the host immune system long-term to ensure prolonged allograft survival. Long-term non-specific immunosuppression can, however, result in life-threatening complications. Thus, efforts have been pursued to explore novel strategies that would allow minimization of maintenance immunosuppression, eventually leading to transplant tolerance. In this scenario, bone marrow-derived mesenchymal stromal cells (MSC), given their unique immunomodulatory properties to skew the balance between regulatory and memory T cells, have emerged as potential candidates for cell-based therapy to promote immune tolerance. Here, we review our initial clinical experience with bone marrow-derived MSC in living-donor kidney transplant recipients and provide an overview of the available results of other clinical programs with MSC in kidney and liver transplantation, highlighting hurdles and success of this innovative cell-based therapy.
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http://dx.doi.org/10.1016/j.humimm.2017.12.008DOI Listing
May 2018

Mesenchymal Stromal Cells for AKI after Cardiac Surgery.

J Am Soc Nephrol 2018 01 11;29(1):7-9. Epub 2017 Dec 11.

IRCCS, Istituto di Ricerche Farmacologiche Mario Negri, Bergamo, Italy.

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http://dx.doi.org/10.1681/ASN.2017111207DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5748931PMC
January 2018

Clinical Translation of Mesenchymal Stromal Cell Therapies in Nephrology.

J Am Soc Nephrol 2018 02 30;29(2):362-375. Epub 2017 Nov 30.

Department of Renal Medicine and Molecular Medicine, IRCCS, Istituto di Ricerche Farmacologiche Mario Negri, Bergamo, Italy.

Mesenchymal stromal cells have emerged as potential candidates for cell-based therapies to modulate the immune response in organ transplantation and repair tissues after acute or chronic injury. Preclinical studies have shown convincingly in rodent models that mesenchymal stromal cells can prolong solid organ graft survival and that they can induce immune tolerance, accelerate recovery from AKI, and promote functional improvement in chronic nephropathies. Multiple complex properties of the cells, including immunomodulatory, anti-inflammatory, and proregenerative effects, seem to contribute. The promising preclinical studies have encouraged investigators to explore the safety, tolerability, and efficacy of mesenchymal stromal cell-based therapy in pilot clinical trials, including those for bone marrow and solid organ transplantation, autoimmune diseases, and tissue and organ repair. Here, we review the available data on culture-expanded mesenchymal stromal cells tested in renal transplantation, AKI, and CKD. We also briefly discuss the relevant issues that must be addressed to ensure rigorous assessment of the safety and efficacy of mesenchymal stromal cell therapies to allow the translation of this research into the practice of clinical nephrology.
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http://dx.doi.org/10.1681/ASN.2017070781DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5791082PMC
February 2018

Mesenchymal stromal cells in renal transplantation: opportunities and challenges.

Nat Rev Nephrol 2016 Apr 8;12(4):241-53. Epub 2016 Feb 8.

IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Clinical Research Center for Rare Diseases "Aldo &Cele Daccò", Via G.B. Camozzi 3, 24020 Ranica, Bergamo, Italy.

Lifelong immunosuppressive therapy is essential to prevent allograft rejection in transplant recipients. Long-term, nonspecific immunosuppression can, however, result in life-threatening complications and fail to prevent chronic graft rejection. Bone marrow (BM)-derived multipotent mesenchymal stromal cells (MSCs) have emerged as a potential candidate for cell-based therapy to modulate the immune response in organ transplantation. These cells can repair tissue after injury and downregulate many of the effector functions of immune cells that participate in the alloimmune response, converting them into regulatory cells. The findings of preclinical and initial clinical studies support the potential tolerance-inducing effects of MSCs and highlight the unanticipated complexity of MSC therapy in kidney transplantation. In animal models of transplantation MSCs promote donor-specific tolerance through the generation of regulatory T cells and antigen-presenting cells. In some settings, however, MSCs can acquire proinflammatory properties and contribute to allograft dysfunction. The available data from small clinical studies suggest that cell infusion is safe and well tolerated by kidney transplant recipients. Ongoing and future trials will provide evidence regarding the long-term safety of MSC therapy and determine the optimum cell source (either autologous or allogeneic) and infusion protocol to achieve operational tolerance in kidney transplant recipients. These studies will also provide additional evidence regarding the risks and benefits of MSC infusion and will hopefully offer definitive answers to the important questions of when, where, how many and which types of MSCs should be infused to fully exploit their immunomodulatory, pro-tolerogenic and tissue-repairing properties.
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http://dx.doi.org/10.1038/nrneph.2016.7DOI Listing
April 2016

Recent advances in immunosuppression and acquired immune tolerance in renal transplants.

Am J Physiol Renal Physiol 2016 Mar 20;310(6):F446-53. Epub 2016 Jan 20.

IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Transplant Research Center "Chiara Cucchi de Alessandri e Gilberto Crespi," Ranica, Bergamo, Italy; IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Clinical Research Center for Rare Diseases "Aldo e Cele Daccò," Ranica, Bergamo, Italy; Unit of Nephrology and Dialysis, Azienda Ospedaliera Papa Giovanni XXIII, Bergamo, Italy; and Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy.

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http://dx.doi.org/10.1152/ajprenal.00312.2015DOI Listing
March 2016

B Cell Reconstitution after Rituximab Treatment in Idiopathic Nephrotic Syndrome.

J Am Soc Nephrol 2016 06 13;27(6):1811-22. Epub 2015 Nov 13.

Division of Nephrology and Dialysis.

The pathogenesis of nephrotic syndrome is unclear. However, the efficacy of rituximab, a B cell-depleting antibody, in nephrotic syndrome suggests a pathogenic role of B cells. In this retrospective study, we determined by flow cytometry levels of B and T cell subpopulations before and after rituximab infusion in 28 pediatric patients with frequently relapsing or steroid-dependent nephrotic syndrome. At baseline, patients had lower median percentages of transitional and mature B cells than age-matched healthy controls (P<0.001). Rituximab induced full depletion of B cells (<1% of lymphocytes). At 1 year, most patients exhibited complete total and mature B cell recovery, whereas memory B cell subsets remained significantly depleted. Total T cell concentration did not change with rituximab, whereas the CD4(+)/CD8(+) T cell ratio tended to increase. Fourteen patients relapsed within 24 months, with a median follow-up of 11.2 months (interquartile range, 8-17.7 months). We observed no difference at baseline between nonrelapsing and relapsing patients in several clinical parameters and cell subset concentrations. Reconstitution of all memory B cell subpopulations, number of immunosuppressive drugs, and dose of tacrolimus during the last 4 months of follow-up were predictive of relapse in univariate Cox regression analysis. However, only delayed reconstitution of switched memory B cells, independent of immunosuppressive treatment, was protective against relapse in multivariate (P<0.01) and receiver operator characteristic (P<0.01 for percentage of lymphocytes; P=0.02 for absolute count) analyses. Evaluation of switched memory B cell recovery after rituximab may be useful for predicting relapse in patients with nephrotic syndrome.
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http://dx.doi.org/10.1681/ASN.2015050523DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4884116PMC
June 2016

Mesenchymal stromal cells to control donor-specific memory T cells in solid organ transplantation.

Curr Opin Organ Transplant 2015 Feb;20(1):79-85

Clinical Research Center for Rare Diseases 'Aldo & Cele Daccò', IRCCS-Istituto di Ricerche Farmacologiche 'Mario Negri', Bergamo, Italy.

Purpose Of Review: Mesenchymal stromal cells (MSCs) represent a promising cell therapy to promote transplant tolerance, as they influence many cells involved in immune response. Herein, we review recent evidence on the ability of MSCs to inhibit antigen-induced memory T cell response in vitro and in preclinical studies as well as immunological studies in kidney transplant recipients highlighting the effects of MSC therapy on memory CD8 T-cell proliferation and function.

Recent Findings: MSCs are able to inhibit in-vitro proliferation and effector functions of memory T cells in response to auto-antigen and allo-antigen stimulation. MSC infusion in animal transplant models resulted in a skew of the balance between regulatory T cells and effector/memory T cells towards a pro-tolerogenic profile. MSC in clinical transplantation is in its infancy and limited numbers of clinical studies have performed immunomonitoring of MSC-treated patients. However, available data support the capability of MSCs to control effector/memory CD8 T-cell proliferation and donor-specific CD8 T-cell function long lasting in kidney transplant setting.

Summary: Recent studies of MSCs in kidney transplantation highlight the anticipated add-on value of the immunomodulatory properties of bone marrow derived MSCs in persistently inhibiting donor-specific effector/memory CD8 T cells, an effect not shared by the current immunosuppressive drugs.
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http://dx.doi.org/10.1097/MOT.0000000000000145DOI Listing
February 2015

Pluripotent stem cells and tolerance induction in organ transplantation.

Curr Opin Organ Transplant 2015 Feb;20(1):86-93

aCentro Anna Maria Astori, Parco Scientifico Tecnologico Kilometro Rosso, IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Bergamo bCentro Ricerche di Medicina Rigenerativa-Fondazione IRCCS Policlinico San Matteo, Pavia cCentro Ricerche Trapianti 'Chiara Cucchi de Alessandri e Gilberto Crespi', IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Bergamo, Italy dFondazione IRCCS-Policlinico San Matteo, Pavia, Italy.

Purpose Of Review: Ongoing research is constantly looking for means to modulate the immune system for long-lasting engraftment of pluripotent stem cells (PSC) during stem cell-based therapies. This study reviews data on in-vitro and in-vivo immunogenicity of embryonic and induced-PSC and describes how their immunological properties can be harnessed for tolerance induction in organ transplantation.

Recent Findings: Although PSC display immunomodulatory properties in vitro, they are capable of eliciting an immune response that leads to cell rejection when transplanted into immune-competent recipients. Nevertheless, long-term acceptance of PSC-derived cells/tissues in an allogeneic environment can be achieved using minimal host conditioning. Protocols for differentiating PSC towards haematopoietic stem cells, thymic epithelial precursors, dendritic cells, regulatory T cells and myeloid-derived suppressor cells are being developed, suggesting the possibility to use PSC-derived immunomodulatory cells to induce tolerance to a solid organ transplant.

Summary: PSC and/or their derivatives possess unique immunological properties that allow for acceptance of PSC-derived tissue with minimal host conditioning. Investigators involved either in regenerative or in transplant medicine must join their efforts with the ultimate aim of using PSC as a source of donor-specific cells that would create a protolerogenic environment to achieve tolerance in solid organ transplantation.
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http://dx.doi.org/10.1097/MOT.0000000000000144DOI Listing
February 2015

Assessment of anti-donor T cell proliferation and cytotoxic T lymphocyte-mediated lympholysis in living donor kidney transplant patients.

Methods Mol Biol 2014 ;1213:355-64

Department of Translational and Regenerative Medicine, Research Block B, 5th Floor, Postgraduate Institute of Medical Education and Research, Sector-12, Chandigarh, 160012, India,

Organ transplant recipients are at risk of allograft rejection, and remain dependent on lifelong immunosuppressive agents, with the attendant risks of infections, cancers, and drug toxicities. Mesenchymal stromal cells (MSCs) have emerged as an alternative to the current pharmacologic immunosuppressive therapy as these cells are immune privileged and possess immunomodulatory properties. However, clinical data are incomplete regarding the efficacy of MSC therapy to control alloimmune response of the transplant recipients. Coordinated efforts should now be directed towards assays for monitoring anti-donor T cell response of MSC-treated patients to establish the pro-tolerogenic potential of MSC-based therapy. Here, we describe two useful tools to monitor MSC-mediated immunomodulation: the assessment of T cell proliferation by carboxyfluorescein succinimidyl ester (CFSE) dilution assay and the evaluation of cytotoxic T lymphocyte (CTL)-mediated lysis of (51)Cr-labeled target cells (cell-mediated lympholysis; CML) following mixed lymphocyte cultures of peripheral blood mononuclear cells (PBMCs) from kidney donors and transplant recipients.
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http://dx.doi.org/10.1007/978-1-4939-1453-1_29DOI Listing
May 2015

Recellularization of well-preserved acellular kidney scaffold using embryonic stem cells.

Tissue Eng Part A 2014 May 29;20(9-10):1486-98. Epub 2014 Jan 29.

1 IRCCS-Istituto di Ricerche Farmacologiche Mario Negri , Bergamo, Italy .

For chronic kidney diseases, there is little chance that the vast majority of world's population will have access to renal replacement therapy with dialysis or transplantation. Tissue engineering would help to address this shortcoming by regeneration of damaged kidney using naturally occurring scaffolds seeded with precursor renal cells. The aims of the present study were to optimize the production of three-dimensional (3D) rat whole-kidney scaffolds by shortening the duration of organ decellularization process using detergents that avoid nonionic compounds, to investigate integrity of extracellular matrix (ECM) structure and to enhance the efficacy of scaffold cellularization using physiological perfusion method. Intact rat kidneys were successfully decellularized after 17 h perfusion with sodium dodecyl sulfate. The whole-kidney scaffolds preserved the 3D architecture of blood vessels, glomeruli, and tubuli as shown by transmission and scanning electron microscopy. Micro-computerized tomography (micro-CT) scan confirmed integrity, patency, and connection of the vascular network. Collagen IV, laminin, and fibronectin staining of decellularized scaffolds were similar to those of native kidney tissues. After infusion of whole-kidney scaffolds with murine embryonic stem (mES) cells through the renal artery, and pressure-controlled perfusion with recirculating cell medium for 24 and 72 h, seeded cells were almost completely retained into the organ and uniformly distributed in the vascular network and glomerular capillaries without major signs of apoptosis. Occasionally, mES cells reached peritubular capillary and tubular compartment. We observed the loss of cell pluripotency and the start of differentiation toward meso-endodermal lineage. Our findings indicate that, with the proposed optimized protocol, rat kidneys can be efficiently decellularized to produce renal ECM scaffolds in a relatively short time, and rapid recellularization of vascular structures and glomeruli. This experimental setup may open the possibility to obtain differentiation of stem cells with long lasting in vitro perfusion.
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http://dx.doi.org/10.1089/ten.TEA.2013.0269DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4011423PMC
May 2014

Mesenchymal stromal cells to promote kidney transplantation tolerance.

Curr Opin Organ Transplant 2014 Feb;19(1):47-53

Department of Clinical Immunology and Transplantation, Azienda Ospedaliera Papa Giovanni XXIII - IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Ricerche Trapianti 'Chiara Cucchi de Alessandri e Gilberto Crespi', Bergamo, Italy.

Purpose Of Review: Cell therapy with mesenchymal stromal cells (MSC) has emerged as a promising tolerance-inducing strategy, as MSC are potent modifiers of immune cells within adaptive as well as innate arm of the immune system. Here, we review recent evidence on both the beneficial and deleterious effect of MSC in experimental models of solid organ transplantation as well as first clinical experiences of MSC therapy in kidney transplant recipients.

Recent Findings: MSC are able to reprogram macrophages toward an anti-inflammatory phenotype capable to regulate antigraft immune response. This interaction is mediated mainly by TNF-α-induced-protein-6. Conversely, MSC also take on a proinflammatory phenotype and actually could worsen graft outcome. MSC in clinical transplantation is in its infancy and nobody so far has attempted to or provided evidence that this cell-based therapy is capable to promote operational tolerance. There are, however, supporting data of the ex-vivo immunoregulatory activity of MSC in treated patients.

Summary: MSC have a great potential as a tolerance-promoting cell therapy. Extensive investigations are still needed to dissect the mechanism(s) of action of MSC, particularly in the setting of a proinflammatory environment, and to establish specific assays for monitoring MSC-treated patients to define the protolerogenic potential of MSC-based therapy in kidney transplantation.
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http://dx.doi.org/10.1097/MOT.0000000000000035DOI Listing
February 2014
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