Periodic Reporting for period 2 - BIRDIE (Bioprinting on-chip microphysiological models of humanized kidney tubulointerstitium)
Période du rapport: 2022-03-01 au 2023-08-31
CKD affects eight hundred and fifty million people, being the 11th leading cause of mortality worldwide.[4] With the progress of CKD, patients with a renal filtration rate below 10% of the normal filtration capacity are diagnosed with an end-stage renal disease (ESRD). At this stage, the patients have to undergo lifesaving dialysis and are placed in a kidney transplant (KT) waiting list. Despite of the advances of dialysis, the filtration functions of the kidneys are not fully replaced and the health of the patients gradually declines. KT is the first choice treatment for ESRD but organ shortage is one limitation. Once a suitable donor organ is identified, the patient undergoes KT where the organ has an average lifespan of 10 years.
Today approximately 90.000 patients are waiting for a suitable donor kidney in Europe and a similar number was reported in US.[5] Immunosuppressive drugs (ISD) have revolutionized the management of KT over the past 30 years by constantly improving the rate of organ survival. However, ISD can promote microbial infections in transplant patients. In particular, viral infections have shown to affect one third of KT patients in the US during the 1995-2002 period.[7]
Kidneys from donors are often infected with cytomegalovirus, BK polyoma virus (BKPyV) and even hepatitis C, and can still be successfully grafted but require strict follow-up and antiviral treatments if available.[8-9] No suitable treatment has been identified so far to treat the BKPyV infection representing now a main challenge for patients currently being addressed by scientists and clinicians. BKPyV reactivation can occur in almost 50% of KT patients which leads to graft loss in 15%-50% of viremic patients.[10] [11]
Renal nephrotoxicity through acute or prolonged exposures to drugs (e.g. nonsteroidal anti-inflammatory drugs), is a recognized problem where patients develop AKI and CKD while being treated for other pathologies. The incidence of drug-induced nephrotoxicity varies up to 26% in adult and up to 16% in pediatric patients.[13] Furthermore, nephrotoxicity is also frequently associated to novel therapeutic treatments where the drugs have shown no toxicity in preclinical studies but prove less safe during clinical trials stages. In some cases, drugs cleared during the clinical trials have been withdrawn from the market at a later stage due to AKI events.
Therefore, In BIRDIE project, we propose to:
* Develop and implement the dual organ-chip-model to investigate BKPyV infection and drug induced AKI. We will use a TissUse (TU, high-tech SME partner of BIRDIE) proprietary dual organ-on-chip (HUMIMIC chip) to better mimic the complexity of the renal environment by including stromal (i.e. fibroblasts) and immune cells (i.e. myeloid dendritic cells), which are thought to participate to early pathophysiology of the BKPyV infection. A “personalized” version of the model will be aimed by applying iPSCs-derived renal progenitor cells;
* Develop strategy allowing bioprinting of renal tubulointerstitium 3D model. Parallel peritubular capillary and proximal tubule will be bioprinted mimicking the cellular and extracellular composition of the native kidney. Microfluidics or pressure based bioprinting will be used to manufacture micron-size tubules (< 70 μm). The interstitial cellular and extracellular matrix compositions will be manufactured with novel bioprinting technique. This model will be initially manufactured with cell lines and once the manufacturing strategy is optimized iPSCs-derived cells will be used;
* Combine the above mentioned objectives to achieve a physiologically relevant bioprinted on-chip 3D model to study viral- and drug-induced AKI. To obtain a further maturation of the bioprinted tissue and to perform long-term cultures with perfusion, the bioprinted model will be combined with the novel on-chip platform.
Today approximately 90.000 patients are waiting for a suitable donor kidney in Europe and a similar number was reported in US.[5] Immunosuppressive drugs (ISD) have revolutionized the management of KT over the past 30 years by constantly improving the rate of organ survival. However, ISD can promote microbial infections in transplant patients. In particular, viral infections have shown to affect one third of KT patients in the US during the 1995-2002 period.[7]
Kidneys from donors are often infected with cytomegalovirus, BK polyoma virus (BKPyV) and even hepatitis C, and can still be successfully grafted but require strict follow-up and antiviral treatments if available.[8-9] No suitable treatment has been identified so far to treat the BKPyV infection representing now a main challenge for patients currently being addressed by scientists and clinicians. BKPyV reactivation can occur in almost 50% of KT patients which leads to graft loss in 15%-50% of viremic patients.[10] [11]
Renal nephrotoxicity through acute or prolonged exposures to drugs (e.g. nonsteroidal anti-inflammatory drugs), is a recognized problem where patients develop AKI and CKD while being treated for other pathologies. The incidence of drug-induced nephrotoxicity varies up to 26% in adult and up to 16% in pediatric patients.[13] Furthermore, nephrotoxicity is also frequently associated to novel therapeutic treatments where the drugs have shown no toxicity in preclinical studies but prove less safe during clinical trials stages. In some cases, drugs cleared during the clinical trials have been withdrawn from the market at a later stage due to AKI events.
Therefore, In BIRDIE project, we propose to:
* Develop and implement the dual organ-chip-model to investigate BKPyV infection and drug induced AKI. We will use a TissUse (TU, high-tech SME partner of BIRDIE) proprietary dual organ-on-chip (HUMIMIC chip) to better mimic the complexity of the renal environment by including stromal (i.e. fibroblasts) and immune cells (i.e. myeloid dendritic cells), which are thought to participate to early pathophysiology of the BKPyV infection. A “personalized” version of the model will be aimed by applying iPSCs-derived renal progenitor cells;
* Develop strategy allowing bioprinting of renal tubulointerstitium 3D model. Parallel peritubular capillary and proximal tubule will be bioprinted mimicking the cellular and extracellular composition of the native kidney. Microfluidics or pressure based bioprinting will be used to manufacture micron-size tubules (< 70 μm). The interstitial cellular and extracellular matrix compositions will be manufactured with novel bioprinting technique. This model will be initially manufactured with cell lines and once the manufacturing strategy is optimized iPSCs-derived cells will be used;
* Combine the above mentioned objectives to achieve a physiologically relevant bioprinted on-chip 3D model to study viral- and drug-induced AKI. To obtain a further maturation of the bioprinted tissue and to perform long-term cultures with perfusion, the bioprinted model will be combined with the novel on-chip platform.
All partners in BIRDIE project have collaborated and exchanged information, knowledge and technology. Nantes Université (NU) has optimized culture conditions of primary renal proximal tubule epithelial cells (hRPTEC) in HUMIMIC chip for development of improved in vitro model. The Biopen technology from FluiCell (FC) was transferred to NU and researchers were trained. The protocols of nanoparticles (virus) delivery to the cells was developed and is being tested. We started to set up the RNAseq and spatial transcriptomic pipeline and acquired data for native renal cells/tissue biopsies (control, BKPyV and T-cell rejection). Moreover, at University of Maastricht (UM) we have optimized hiPSC differentiation to obtain metanephric mesenchyme and ureteric bud progenitors for 3D bioprinting. The Biopixlar bioprinter from FC was transferred and initial testing’s were performed. We are working on developing new coating strategies to allow further support for cells. We transferred a second human induced pluripotent stem cell line from TU to UM and we have initiated the validation of the differentiation protocol with this line. Moreover, TU optimized the cryopreservation of organoids and tested the shipment to NU. FC redesigned the optical path of the bioprinter and begun to develop means of integrating the Biopen and Biopixlar fluidic heads into the same platform.
The in vitro models have the potential to help developing new therapies for patients receiving a donor organ. With the combined effect of the immunosuppressive drugs and the presence of BKPyV virus, the treatment for these patients should ensure that viral infection is cleared or controlled. The aimed in BIRDIE project in vitro models will be essential to test new therapies administered to patients (e.g. during drug development) or ultimately being able to generate patient-specific in vitro models (derived for iPSCs generated from patient own cells) allowing personalized medicine approaches. The creation of more human-relevant data might lead to an improved understanding of kidney disease and its treatment, and the reduction in vivo trials.