ICF: Hollow-fiber bioreactor technology to explore mechanisms and delivery of cellular therapy during machine perfusion of donated human kidneys
Lead Research Organisation:
Newcastle University
Department Name: Translational and Clinical Res Institute
Abstract
Kidneys have many functions, including cleaning the blood and removing waste. If the kidneys fail a kidney transplant is almost always the best treatment. Most kidney transplants in the UK come from deceased donors. The process of removing, transporting, and transplanting a kidney takes time and causes damage. Some kidneys which are working well in the potential organ donor cannot tolerate this damage, and therefore are not used for transplant. This lack of suitable kidneys leads to long waiting times for people with kidney failure. Some people who need a kidney transplant never get one
Many treatments have been tried to limit the damaging effects of removing, transporting, and transplanting these kidneys. One option involves a machine which pumps fluid around the kidney whilst it is outside of the body. This is called 'machine perfusion'. It allows the delivery of other treatments directly to a kidney
Our research group has shown that a cell therapy using Multipotent Adult Progenitor Cells or MAPC® cells can be delivered to a kidney during machine perfusion. The MAPC cells, taken from human bone marrow and grown in a laboratory, were delivered into the kidney through its artery. They appeared to improve the quality of the kidneys. However, there are two main barriers preventing us using these kidneys for transplant. Firstly, it is not well understood exactly how the MAPC cells work in this setting. Secondly, the best method for delivering the MAPC cells to the kidney is not known. This research project aims to remove those barriers
To achieve this aim, I will use a new method of delivering this cell treatment. Instead of putting the MAPC cells into the kidney, the MAPC cells will be maintained in a plastic container called a 'bioreactor'. I have spent 9 months finding the best conditions to grow MAPC cells in this bioreactor. I have also used fluid from previous kidney machine perfusions and shown that this 'activates' MAPC cells in the bioreactor, which will enable them to have an impact on the kidney
Moving this project forward will involve performing machine perfusion on pairs of human kidneys. These kidneys will have been donated, but then deemed unsuitable for transplantation. We can use this valuable gift in this approved research project where the next of kin gives consent. This set of kidneys will not be transplanted after our experiments
We will include a bioreactor of MAPC cells as part of the machine perfusion device. The MAPC cells and the kidney can communicate, meaning the MAPC cells can treat the kidney. However, the MAPC cells and the kidney are in separate compartments. This gives the unique opportunity to study the interaction between the kidney and the MAPC cells in a controlled way. Importantly, we can see what effects the kidney has on the MAPC cells. Changing the properties of the bioreactor will allow me to modify how the two communicate and interact
This is a unique type of experiment. I hope that this will answer several important questions: How are MAPC cells activated? How do MAPC cells in a bioreactor affect a human kidney? How do they have this effect? What types of interaction are needed for them to have their effect? Answering these questions will bring us closer to transplanting kidneys using this treatment. This could increase the number of kidneys available for transplant, reducing waiting times, and giving hope to patients on the wait list
The potential implications of this research are also more wide reaching. The processes which injure donated organs also happen in common health problems like heart attacks and strokes. For example, there are currently clinical trials looking at MAPC cells as a treatment for strokes. New insights into how these cell therapies are activated, how they have their effect, and how they can be delivered could be applicable in a wide range of settings. Findings from this project could therefore lead to improved treatments for a much wider range of diseases
Many treatments have been tried to limit the damaging effects of removing, transporting, and transplanting these kidneys. One option involves a machine which pumps fluid around the kidney whilst it is outside of the body. This is called 'machine perfusion'. It allows the delivery of other treatments directly to a kidney
Our research group has shown that a cell therapy using Multipotent Adult Progenitor Cells or MAPC® cells can be delivered to a kidney during machine perfusion. The MAPC cells, taken from human bone marrow and grown in a laboratory, were delivered into the kidney through its artery. They appeared to improve the quality of the kidneys. However, there are two main barriers preventing us using these kidneys for transplant. Firstly, it is not well understood exactly how the MAPC cells work in this setting. Secondly, the best method for delivering the MAPC cells to the kidney is not known. This research project aims to remove those barriers
To achieve this aim, I will use a new method of delivering this cell treatment. Instead of putting the MAPC cells into the kidney, the MAPC cells will be maintained in a plastic container called a 'bioreactor'. I have spent 9 months finding the best conditions to grow MAPC cells in this bioreactor. I have also used fluid from previous kidney machine perfusions and shown that this 'activates' MAPC cells in the bioreactor, which will enable them to have an impact on the kidney
Moving this project forward will involve performing machine perfusion on pairs of human kidneys. These kidneys will have been donated, but then deemed unsuitable for transplantation. We can use this valuable gift in this approved research project where the next of kin gives consent. This set of kidneys will not be transplanted after our experiments
We will include a bioreactor of MAPC cells as part of the machine perfusion device. The MAPC cells and the kidney can communicate, meaning the MAPC cells can treat the kidney. However, the MAPC cells and the kidney are in separate compartments. This gives the unique opportunity to study the interaction between the kidney and the MAPC cells in a controlled way. Importantly, we can see what effects the kidney has on the MAPC cells. Changing the properties of the bioreactor will allow me to modify how the two communicate and interact
This is a unique type of experiment. I hope that this will answer several important questions: How are MAPC cells activated? How do MAPC cells in a bioreactor affect a human kidney? How do they have this effect? What types of interaction are needed for them to have their effect? Answering these questions will bring us closer to transplanting kidneys using this treatment. This could increase the number of kidneys available for transplant, reducing waiting times, and giving hope to patients on the wait list
The potential implications of this research are also more wide reaching. The processes which injure donated organs also happen in common health problems like heart attacks and strokes. For example, there are currently clinical trials looking at MAPC cells as a treatment for strokes. New insights into how these cell therapies are activated, how they have their effect, and how they can be delivered could be applicable in a wide range of settings. Findings from this project could therefore lead to improved treatments for a much wider range of diseases
Technical Summary
Kidney transplantation is the optimal renal replacement therapy, yet a shortage of high-quality organs limits the number of transplants performed. Normothermic machine perfusion (NMP) is a technique where an organ is pumped ex situ with an oxygenated fluid (perfusate). We have shown Multipotent Adult Progenitor Cells (MAPC® cells), an MSC-like cellular therapy, have therapeutic effects in preclinical renal NMP studies when delivered directly into a renal artery. However, the mechanism of action and optimal delivery method is poorly understood; both are barriers to translation.
This project aims to explore MAPC cell licencing (activation) and mechanisms of action by establishing a novel method of MAPC cell delivery during NMP, using hollow-fiber bioreactor technology. MAPC cells can be retained outside of hollow-fibers, interacting with perfusate flowing within fibers, allowing bi-directional interaction between damaged kidneys and MAPC cells.
NMP will be performed on human kidney pairs which have been declined for transplant; one kidney will be randomised to treatment, and the other to control. Hollow-fiber pore size can be manipulated to control signalling mechanisms between the MAPC cells and kidney; for example, 200nm pores would permit extracellular vesicles to pass, whereas restricting to 70kDa will only allow soluble signalling molecules. As the MAPC cells are retained in the bioreactor, RNA can be extracted following NMP; I will use RNA-seq to assess transcriptomic changes in 'licenced' MAPC cells. Therapeutic effect will be assessed using validated outputs including urine production and perfusate/urine injury markers. This dynamic system with controlled signalling between cells and an isolated human organ gives a unique opportunity for mechanistic assessment. This will facilitate dissecting the mechanism of cell licencing and therapeutic effect, bringing us closer to translating NMP MAPC cell therapy, and has ramifications for researchers investigating cellular therapies in a wide variety of clinical settings
This project aims to explore MAPC cell licencing (activation) and mechanisms of action by establishing a novel method of MAPC cell delivery during NMP, using hollow-fiber bioreactor technology. MAPC cells can be retained outside of hollow-fibers, interacting with perfusate flowing within fibers, allowing bi-directional interaction between damaged kidneys and MAPC cells.
NMP will be performed on human kidney pairs which have been declined for transplant; one kidney will be randomised to treatment, and the other to control. Hollow-fiber pore size can be manipulated to control signalling mechanisms between the MAPC cells and kidney; for example, 200nm pores would permit extracellular vesicles to pass, whereas restricting to 70kDa will only allow soluble signalling molecules. As the MAPC cells are retained in the bioreactor, RNA can be extracted following NMP; I will use RNA-seq to assess transcriptomic changes in 'licenced' MAPC cells. Therapeutic effect will be assessed using validated outputs including urine production and perfusate/urine injury markers. This dynamic system with controlled signalling between cells and an isolated human organ gives a unique opportunity for mechanistic assessment. This will facilitate dissecting the mechanism of cell licencing and therapeutic effect, bringing us closer to translating NMP MAPC cell therapy, and has ramifications for researchers investigating cellular therapies in a wide variety of clinical settings
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| Samuel Tingle (Principal Investigator / Fellow) |