Regenerative medicine applications of cholangiocyte organoids
Lead Research Organisation:
University of Cambridge
Department Name: Medicine
Abstract
Transplantation remains the only treatment for all end-stage diseases; but it is limited by organ availability. Regenerative medicine could provide a therapeutic alternative, using cells or lab-grown artificial tissue to regenerate or replace damaged organs. This approach is particularly relevant in bile duct diseases, which are a leading cause for liver transplantation in children and adults with no other treatment.
The bile ducts are tubes transferring a toxic digestive fluid, called bile, from the liver to the intestine. In disease, bile ducts break down and bile damages surrounding tissue causing liver failure. To repair this damage, I grew human duct cells in the lab, in 3D structures called organoids. I then made artificial ducts from my organoids, transplanted them in mice and showed their therapeutic potential.
My goal is to progress regenerative medicine using organoids to first-in-man studies. I will first focus on biliary disease, but the technology and know-how developed will be highly transferable to other organs. To achieve this, I will address three main challenges.
1. Optimise organoids grown in the lab for human transplantation
Transplanted cells should match the tissue they are replacing and meet regulatory standards. I will collaborate with Bilitech LTD and the UK Cell and Gene Therapy Catapult (CGTC) to generate organoids complying with requirements for human transplantation. I will compare these organoids to human bile ducts with a high-resolution method called Single Cell RNA sequencing and use this information to further improve my cells and minimize differences.
2. Develop organoid transplantation techniques tailored to different disease indications
Certain diseases may affect too many and too small bile ducts to be amenable to surgical correction with artificial tissue. However, these small branches may be reached and regenerated by organoids injected directly in the ducts, providing a valuable therapeutic alternative to lab-grown tissue. To establish the safety and efficacy of this approach, I will attempt to rescue small animal models of widespread duct damage using organoid injection.
3. Validate the safety and efficacy of my organoids in large animal models and ex-vivo human organs
Organoids can't be transplanted in human organs outside clinical trials, and large animal experiments are a pre-requisite for such studies. In the context of a UKRI feasibility award with Bilitech LTD and CGTC, I have upscaled my artificial bile ducts from mouse to human size. I will transplant these artificial ducts in adult pigs and follow up the animals for 6 months to demonstrate safety and efficacy.
In parallel, I will explore the cells' capacity to engraft in human organs using a new system I developed. I will capture liver grafts not used for transplantation, maintain them outside the body (ex-vivo) by circulating oxygenated blood through the liver, and inject my organoids in these organs to repair their bile ducts.
This work will advance regenerative medicine using organoids from proof-of-principle to first-in-human studies, which is in direct alignment with the UK strategy on Regenerative Medicine (Regenerative Medicine Expert Group report). It will address a pressing clinical need; provide an alternative to liver transplantation, which is the only treatment for bile ducts disorders; and reduce pressure on the transplantation program benefiting all other liver diseases. It will generate transferrable outputs impacting on broader fields, such as tissue engineering, organoid biology and bioinformatics, which can instruct my future research. The collaboration with Bilitech LTD and CGTC will forge ties between academia and industry. The expertise and technology developed will be applicable to other tissues and organs and will make a significant contribution to UK regenerative and medicines manufacturing know-how.
The bile ducts are tubes transferring a toxic digestive fluid, called bile, from the liver to the intestine. In disease, bile ducts break down and bile damages surrounding tissue causing liver failure. To repair this damage, I grew human duct cells in the lab, in 3D structures called organoids. I then made artificial ducts from my organoids, transplanted them in mice and showed their therapeutic potential.
My goal is to progress regenerative medicine using organoids to first-in-man studies. I will first focus on biliary disease, but the technology and know-how developed will be highly transferable to other organs. To achieve this, I will address three main challenges.
1. Optimise organoids grown in the lab for human transplantation
Transplanted cells should match the tissue they are replacing and meet regulatory standards. I will collaborate with Bilitech LTD and the UK Cell and Gene Therapy Catapult (CGTC) to generate organoids complying with requirements for human transplantation. I will compare these organoids to human bile ducts with a high-resolution method called Single Cell RNA sequencing and use this information to further improve my cells and minimize differences.
2. Develop organoid transplantation techniques tailored to different disease indications
Certain diseases may affect too many and too small bile ducts to be amenable to surgical correction with artificial tissue. However, these small branches may be reached and regenerated by organoids injected directly in the ducts, providing a valuable therapeutic alternative to lab-grown tissue. To establish the safety and efficacy of this approach, I will attempt to rescue small animal models of widespread duct damage using organoid injection.
3. Validate the safety and efficacy of my organoids in large animal models and ex-vivo human organs
Organoids can't be transplanted in human organs outside clinical trials, and large animal experiments are a pre-requisite for such studies. In the context of a UKRI feasibility award with Bilitech LTD and CGTC, I have upscaled my artificial bile ducts from mouse to human size. I will transplant these artificial ducts in adult pigs and follow up the animals for 6 months to demonstrate safety and efficacy.
In parallel, I will explore the cells' capacity to engraft in human organs using a new system I developed. I will capture liver grafts not used for transplantation, maintain them outside the body (ex-vivo) by circulating oxygenated blood through the liver, and inject my organoids in these organs to repair their bile ducts.
This work will advance regenerative medicine using organoids from proof-of-principle to first-in-human studies, which is in direct alignment with the UK strategy on Regenerative Medicine (Regenerative Medicine Expert Group report). It will address a pressing clinical need; provide an alternative to liver transplantation, which is the only treatment for bile ducts disorders; and reduce pressure on the transplantation program benefiting all other liver diseases. It will generate transferrable outputs impacting on broader fields, such as tissue engineering, organoid biology and bioinformatics, which can instruct my future research. The collaboration with Bilitech LTD and CGTC will forge ties between academia and industry. The expertise and technology developed will be applicable to other tissues and organs and will make a significant contribution to UK regenerative and medicines manufacturing know-how.
People |
ORCID iD |
Fotios Sampaziotis (Principal Investigator / Fellow) |
Publications
Bargehr J
(2021)
Cardiovascular ACE2 receptor expression in patients undergoing heart transplantation.
in ESC heart failure
Brevini T
(2023)
FXR inhibition may protect from SARS-CoV-2 infection by reducing ACE2.
in Nature
Jalan-Sakrikar N
(2023)
Organoids and regenerative hepatology.
in Hepatology (Baltimore, Md.)
John BV
(2023)
Ursodeoxycholic acid is associated with a reduction in SARS-CoV-2 infection and reduced severity of COVID-19 in patients with cirrhosis.
in Journal of internal medicine
Meng B
(2022)
Altered TMPRSS2 usage by SARS-CoV-2 Omicron impacts infectivity and fusogenicity.
in Nature
Wesley BT
(2022)
Single-cell atlas of human liver development reveals pathways directing hepatic cell fates.
in Nature cell biology
Description | A novel agent which is already approved for clinical use and can be repurposed to prevent COVID-19 infection. Importantly, this drug is not prone to viral resistance and can be administered to high-risk groups where vaccines don't work |
Exploitation Route | The findings have already given rise to clinical studies exploring the impact of this drug on patients. Furthermore, clinical trials using this agent are being considered |
Sectors | Healthcare |
URL | https://www.nature.com/articles/s41586-022-05594-0 |
Description | Our study identified a novel agent that could be repurposed against COVID-19. This set the foundation for further clinical studies and the drug is being considered for a clinical trial which could have an immediate impact on patients. |
First Year Of Impact | 2023 |
Sector | Healthcare |
Impact Types | Societal |
Description | Evelyn Trust Project Grant |
Amount | £188,713 (GBP) |
Funding ID | 22/58 |
Organisation | The Evelyn Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 10/2022 |
End | 09/2025 |
Description | MRC Equip - World Class Labs award 2022/23 |
Amount | £240 (GBP) |
Funding ID | MC_PC_MR/X01228X/1 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 12/2022 |
End | 03/2023 |
Description | Theme-based Research Scheme |
Amount | HK$56,127,000 (HKD) |
Organisation | Research Grants Council Hong Kong |
Sector | Public |
Country | China |
Start | 01/2022 |
End | 12/2027 |
Title | Method for injection of cholangiocyte organoids in human livers |
Description | Injection of cholangiocyte organoids through the common mile duct using a catheter under radiological guidance |
Type Of Material | Technology assay or reagent |
Year Produced | 2021 |
Provided To Others? | Yes |
Impact | Advancing regenerative medicine |
URL | https://science.sciencemag.org/content/371/6531/839 |
Title | Use of human organs perfused ex-situ to model disease and test novel therapeutic approaches |
Description | Human organs are maintained outside the body in near physiological conditions by perfusing them with warm oxygenated blood. Therapeutic agents or pathogens can be administered in the circulating blood or directly in the organ and their effects can be studied. This is the closest platform to human studies to date |
Type Of Material | Model of mechanisms or symptoms - human |
Year Produced | 2023 |
Provided To Others? | Yes |
Impact | We used human lungs and livers perfused ex-situ to: 1. Test a novel agent and its impact on the expression of the SARS-CoV-2 receptor ACE2 on a whole organ level. The same approach could be used to test any therapeutic agents on human organs 2. Model SARS-CoV-2 infection in human organs perfused ex-situ. The same approach could be used to model a variety of other diseases, including infectious diseases |
URL | https://www.nature.com/articles/s41586-022-05594-0 |
Title | scRNAseq of primary cholangiocytes and cholangiocyte organoids (human) |
Description | scRNAseq dataset |
Type Of Material | Database/Collection of data |
Year Produced | 2021 |
Provided To Others? | Yes |
Impact | Publication (Sampaziotis et al, Science, 2021) |
URL | https://science.sciencemag.org/content/371/6531/839 |
Description | Collaboration with Andrew Fisher (University of Newcastle) |
Organisation | University of Newcastle |
Country | Australia |
Sector | Academic/University |
PI Contribution | Isolation of lung tissue and infection with SARS-CoV-2 Design of experiment Provision of UDCA and reagents to be added in the perfusate |
Collaborator Contribution | Provision and ex-vivo perfusion of human lungs Also contributed to designing the experiment |
Impact | Multidisciplinary Transplant surgery, Infectious Diseases 2 papers 1 in revision, 1 published (Meng et al., NAture, 2022) |
Start Year | 2021 |
Description | Collaboration with Andrew Owen |
Organisation | University of Liverpool |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Identified a novel agent against COVID19 to test on animals. Provided the agent to be tested and discovered the mechanism |
Collaborator Contribution | Performed animal experiments |
Impact | Multidisciplinary collaboration resulting in a publication https://www.nature.com/articles/s41586-022-05594-0 |
Start Year | 2022 |
Description | Collaboration with Binu John to explore the impact of UDCA against SARS-CoV-2 on patient cohorts |
Organisation | University of Miami |
Country | United States |
Sector | Academic/University |
PI Contribution | Discovered a new agent that could be repurposed against COVID19 |
Collaborator Contribution | Performed a retrospective clinical study |
Impact | First manuscript published (https://www.nature.com/articles/s41586-022-05594-0) Ongoing work with second manuscript in submission |
Start Year | 2022 |
Description | Collaboration with CITIID (Steve Baker, Gordon Dougan, Ravi Gupta, Nicholas Matheson, Paul Lehner) |
Organisation | University of Cambridge |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provision of cholangiocyte organoids for infection with SARS-CoV-2 |
Collaborator Contribution | Provision of SARS-CoV-2 virus, primers to measure virus, antibodies, fluorescent spike protein, HEK cell lines, training in infection |
Impact | 2 papers 1 submitted - in revision 1 published in Nature (see publications, Meng et al, Nature, 2022) |
Start Year | 2021 |
Description | Collaboration with Michael Trauner (University of Vienna) |
Organisation | University of Vienna |
Country | Austria |
Sector | Academic/University |
PI Contribution | Training members of the Trauner Lab to grow organoids Contribution to in vitro testing of new drugs for cholangiopathies on cholangiocyte organoids |
Collaborator Contribution | In vivo experiments in mice |
Impact | Part of a multidisciplinary collaboration Outputs Short term EASL fellowship for Claudia Fuchs to visit Cambridge (Trauner lab) Paper submission (in revision) |
Start Year | 2021 |
Title | Cell Transplantation |
Description | A novel method to inject cells in human organs perfused ex-situ |
IP Reference | PCT/EP2022/053910 |
Protection | Patent / Patent application |
Year Protection Granted | 2022 |
Licensed | Commercial In Confidence |
Impact | Using this method we have managed to successfully inject cells and repair human livers perfused ex-situ |
Title | METHODS OF EXPANDING CHOLANGIOCYTES |
Description | This invention relates to the expansion of primary cholangiocytes in the form of cholangiocyte organoids (COs) using culture conditions in which canonical Wnt signalling is inhibited and non-canonical Wnt/PCP signalling is potentiated. Methods of expanding primary cholangiocytes, expanded populations of cholangiocytes and medical applications of expanded cholangiocytes are provided. |
IP Reference | CA3067759 |
Protection | Patent / Patent application |
Year Protection Granted | 2018 |
Licensed | Commercial In Confidence |
Impact | Cells generated with this method have been shown to successfully regenerate human livers perfused ex-situ |
Title | Treatment of viral infection |
Description | The present inventors have discovered that reducing the expression or activity of the farnesoid X receptor (FXR) leads to the downregulation of angiotensin converting enzyme 2 (ACE2) expression in cells. Reducing FXR expression or activity in cells or tissues may therefore be useful in treating ACE2 mediated diseases, and reducing the susceptibility of individuals to such diseases. Reducing FXR expression or activity is shown herein to reduce or prevent the entry of ACE2 binding viruses into the cells. In particular, the expression of ACE2 is reduced at sites that interact with the ACE2 binding virus. This may be useful in treating ACE2 mediated infection and reducing the susceptibility of individuals to ACE2 mediated viral infection. |
IP Reference | 21386031.5 |
Protection | Patent / Patent application |
Year Protection Granted | 2022 |
Licensed | Commercial In Confidence |
Impact | Identified agent that could be used to prevent COVID19 in high risk groups |
Description | Broadcast for the CUH website |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Media (as a channel to the public) |
Results and Impact | Interview for the CUH website outlining our work on repurposing UDCA as a novel agent to prevent COVID19 in high risk groups |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.cuh.nhs.uk/news/drug-to-treat-liver-disease-could-prevent-covid/ |
Description | Interview for National News |
Form Of Engagement Activity | A broadcast e.g. TV/radio/film/podcast (other than news/press) |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Media (as a channel to the public) |
Results and Impact | BBC East - TV coverage of our most recent work on repurposing the clinically approved drug UDCA to prevent COVID19 in high risk groups |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.youtube.com/watch?v=x03Q_bIYBK0 |
Description | Patient Group Workshop - Ask the experts (Support PSC UK group) |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Explaining our research and future outlooks to PSC patients |
Year(s) Of Engagement Activity | 2021 |
URL | https://www.youtube.com/watch?v=QGTVCONg_Ck |
Description | Podcast |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Podcast outlining the use of organoids for organ regeneration |
Year(s) Of Engagement Activity | 2021 |
URL | https://podcasts.apple.com/us/podcast/lab-grown-organoids-and-regenerative-medicine-with/id135807974... |
Description | TEDx Talk in the University of West Attica |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited TEDx talk entitled Tissue Regeneration - A ticket to improving life? |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.ted.com/tedx/events/37361 |