*PDOS are Patient Derived Organoids
February 28th is Rare Disease Day. In leap years, the day is February 29th, the rarest day of the year. “Rare Disease Day is the globally coordinated movement on rare diseases, working towards equity in social opportunity, healthcare and access to diagnosis and therapies for people living with a rare disease.”
What is meant by FAP and MAP?
Familial Adenomatous Polyposis (FAP) and MUTYH-associated polyposis (MAP) are rare, inherited genetic conditions which predispose to a very high risk of bowel cancer. Patients develop multiple adenomatous polyps. These are small growths on the inner lining of the colon (large intestine), and other areas of the intestinal tract.
Polyps can acquire genetic mutations that disrupt the functioning of the normal cell machinery and cause them to grow quickly, in a dysregulated manner. These polyps are on their way to becoming malignant.
The average age for polyps to develop in patients with FAP is the mid-teens, with multiple polyps developing by the age of 35. MAP may be less severe than FAP with fewer polyps developing and later. The diagnosis is made by analysing the genetic code of the patients to look for specific mutations in the DNA that indicate why the instructions for normal cell growth have been corrupted.
Treatment is by preventative colectomy (removal of some, or all the colon) and regular endoscopic examination of the upper intestines. Endoscopy involves the insertion of a long, thin, flexible tube that has a tiny video camera at the end so that clinicians can closely observe the lining of the intestine as the tube is fed through. Further surgery may be required if polyps form in the duodenum (duodenectomy). Very little is known about the causes of duodenal polyps and how this differs from colorectal disease in FAP and MAP. Research into this area is a high priority as there are high economic and quality of life costs associated with both diseases.
The Inherited Tumour Syndromes Research (ITSR) group at Cardiff University has been developing 3D organoid cell models to represent duodenal malignancy in FAP and MAP patients. In a joint project with Cellesce, these models have been developed further, for use in an industrial environment, to facilitate drug discovery.
Organoids are 3D cell structures that are miniaturised versions of the tissue from which they originated. Organoids derived from intestinal biopsy material from FAP and MAP patients, replicate the biology and appearance of duodenal adenomas or of normal tissue, depending on where the biopsy was taken from. They are used as models for research into the causes and effects of the diseases and as a platform for pre-clinical testing of preventative treatments.
The project succeeded in obtaining living duodenal tissues donated by affected patients. Duodenal adenoma and normal 3D organoid lines from the same patients (for comparison), were successfully derived and expanded.
Duodenal adenoma organoids derived from biopsy tissue from a FAP patient
The ideal treatment for FAP and MAP patients would be to correct or neutralise the effects of the faulty instructions in these patients. This would prevent, or at least delay the growth of polyps and therefore disease progression and cause no unwanted side-effects. To date, no suitable drugs have been discovered that fulfil these needs.
Work is ongoing to find novel treatments and to explore the use of existing drugs that have already been approved for clinical use. This is called drug repurposing. Since safety in humans has already been demonstrated, the drug development timeline and the research and development costs are significantly reduced.
An example of this is a drug called guselkumab (Tremfya), that is used in the treatment of plaque psoriasis. It works by blocking inflammatory and immune responses. Janssen Pharmaceuticals (part of Johnson and Johnson) are performing clinical trials using this drug to treat FAP patients. Additional clinical trials involve the repurposing of icosapent ethyl used to prevent heart attacks (GLW Pharma), sirolimus (Emtora Biosciences), used after renal transplants to prevent rejection of the new kidney and the drugs chosen for the study below.
Pilot drug-treatment study
Sulindac (Merck): a non-steroidal anti-inflammatory
Erlotinib (Astellas Pharma U.S.): a tyrosine kinase inhibitor that slows the growth of cancer cells with specific proteins (“EGFR”) on their cell surface.
To show if the effect of these drugs on 3D organoids mirrors the patient response, duodenal adenoma and the corresponding normal 3D organoid lines will be treated with Sulindac and Erlotinib alone and in combination. If successful, this will demonstrate that 3D duodenal adenoma and normal organoids derived from FAP/MAP patients are a suitable platform on which to test novel compounds and drug combinations for patients with these conditions. Findings from such studies may also be relevant to sporadic intestinal cancer therapy.
At Cellesce we have scaled up the expansion and manufacture of FAP/MAP 3D organoid lines using our unique bioprocessing technology. This will enable them to be used in innovative research and in high-throughput screens by pharmaceutical companies to facilitate drug discovery and development.
The charity Bowel Cancer West supported the initial work to derive and culture FAP and MAP 3D organoid models. Further development of the models was jointly supported by Cellesce and the Clinical Innovation Accelerator through Accelerate, a programme part-funded by the European Regional Development Fund.
None of this work could have taken place without the support and hard work of administrative, research and clinical staff in NHS hospitals, Cardiff University and Cellesce. Neither would it have been possible without the generous donation of biopsy tissue from patients with FAP and MAP, for which we extend our thanks.
The United Nations General Assembly declared 11 February as the International Day of Women and Girls in Science in 2015. This was to bring attention to the fact that women and girls continue to be excluded from science education and are therefore under-represented in university courses and occupations.
The following is paraphrased from the United Nations website:
“Women continue to be under-represented in science, technology, engineering and mathematics (STEM) all over the world. On 20th December 2013, the General Assembly adopted a resolution in which it recognised that full and equal access to and participation in science, technology and innovation for women and girls of all ages is imperative for achieving gender equality and the empowerment of women and girls.”
In the UK, there is no barrier to women participating in science subjects, but only 35% of STEM students are women, demonstrating that even where there is opportunity, there is an unwillingness to consider training and employment in these areas. This is despite the fact that core STEM employment opportunities continue to increase more in comparison to other areas.
Figure taken from “Women in Stem” and based on UCAS data from HESA and findings from WISE campaigns.
There is a national shortage of STEM professionals that needs to be addressed by encouraging the participation of underrepresented groups.
“To stay competitive globally, the nation needs the talent and creative ability of all of its people—both women and men. But women currently are a smaller part of the science and engineering workforce—in industry and in our nation’s colleges and universities.”
Quote taken from the National Academy of Sciences.
Cellesce, was co-founded by Professor Marianne Ellis, an expert in the design of bioprocesses for scalable bioreactors for cell expansion, She is currently Head of Department of Chemical Engineering at Bath University.
At Cellesce, there are 13 full-time members of staff. 9 of these are women (62%).
Here are some soundbites from a few of our scientists, explaining their role at Cellesce and what inspired them to choose science as a career.
Dr. Victoria Marsh-Durban, CEO
“I have overall responsibility and accountability for the day-to-day running of Cellesce as a business. I now work on the business/commercial side of the company but I trained first as a scientist and hold a Ph.D. in Cancer Genetics. I first became interested in science at the age of 7, when I was given the “Usborne Science Encyclopaedia” as a gift – it instantly became my favourite book, particularly the section on biology and the human body! Later, I became fascinated with proteins, and their amazing diversity, function and roles in human health and disease, which led to me studying Biochemistry. I’ve also always enjoyed practical tasks and making/creating things with my hands, so science was a perfect outlet for that too.”
Dr. Carly Bunston, Research Scientist
As a Research Scientist at Cellesce my role involves the optimisation and biological validation of our bioreactor process for the expansion of new patient-derived organoids lines. My interest in cancer research sparked my desire to become a scientist, with the hope that my work will in some way help towards improving patient outcomes.
Elizabeth Fraser, Alliance Manager
I worked for many years in academic labs in the field of Cancer Research. Now, I apply that experience to managing research projects within the company. I also develop relationships with external parties to complement Cellesce’s expertise and help our business to flourish.
My father was an Organic Chemist in the pharmaceutical industry, so he was the person who encouraged me to have an interest in science (although I preferred biology to chemistry)! I’ve always loved crafting and making things and the practicality and dexterity needed for that is very applicable to lab work.
Dr Jessica Pinheiro de Lucena-Thomas, Bioprocess Engineer
I operate and monitor Cellesce’s bioreactors and am currently working on the development of our next generation technology. My first degree was in Chemical Engineering in Brazil. I moved to the UK in 2016 and studied for my Ph.D at the University of Bath.
I was inspired to be an engineer following a visit to the paint manufacturing plant where my father used to work. I loved the massive machinery, pipes and reactors. Today, I am working on a much smaller scale (with organoids), which presents its own challenges. It’s very rewarding to be able to apply my engineering expertise to the development of an enabling technology that will facilitate success in drug discovery and ultimately improve people’s lives.
Harman Chaggar, Manufacturing Technician
As a Manufacturing Technician, I assist in the development of new protocols and processes. Once established, I must closely follow the Standard Operating Procedures of the products being manufactured. This ensures that the organoids that we produce meet the necessary specifications and are consistent and reproducible. I was drawn to science in High School when I learned how advances in technology can be used to increase our understanding of human biology and the mechanisms of disease. With this knowledge comes the ability to look for effective treatments.
What we do at Cellesce:
This animation gives a brief overview of our company.
We have a patented bioprocessing technology that enables Cellesce to generate high quality organoids at scale for the pharma and biotech industries as a more patient-centric model for drug discovery and development.
What are Patient-Derived Organoids (PDOs)? The term OrganOID implies that a PDO resembles an organ; hence another name, “mini-organ”. Unlike organ donation, these miniaturised versions can be grown in the lab from a tiny piece of biopsy tissue from a living donor. Specialised cells within the biopsy, known as adult stem cells, can divide and form new cell types by a process called differentiation. These cells self-assemble in three-dimensions so that they structurally replicate the organ that the tissue was sampled from and faithfully represent the human biology of that individual patient.
The images below were taken using a specialised microscope. The PDOs were stained with blue dye, to show up DNA in individual cells and with yellow dye to show the outline of the lumen. (The equivalent of the gut cavity.) A whole 3D PDO is shown on the left and on the right you can see a slice through the middle).
The PDOs are fully characterised and can be used in research, in areas such as infectious diseases, genetic disorders, toxicology and cancers. A particularly useful application is in drug development, for testing anti-cancer compounds for example.
Drugs that have a detrimental effect on the PDOs that represent a particular cancer sub-type, are likely to be an effective treatment for the same type of cancer in the actual patients themselves.
Using PDO lines from multiple patients to screen for potential treatments, including people from different racial backgrounds and ethnic minority groups, will improve efficiency in drug discovery. Early, accurate prediction of success in the later stages of drug development will reduce the failure rate and associated costs. This will increase the choice of targeted drugs available for clinicians. Ultimately, this will improve survival rates and the quality of life for patients.
10 interesting facts about PDOs
- Culture of Patient Derived Organoids (PDOs)
PDOs can be grown from healthy as well as diseased tissue. PDOs from cancer tissue are often the easiest to culture and grow because the nature of the disease is that cancer cells grow quickly and in a deregulated manner. Scientists must find the optimum culture conditions to allow the PDOs to develop without affecting their biology, which can be more difficult in ‘normal’ tissue, but not impossible.
- The first PDOs to be established were from the colon, creating “mini-guts”
The first detailed report showing that PDOs could be established from human biopsy tissue was from Hans Clevers’ lab in the Netherlands (Sato et al., 2009). Colon biopsy tissue was processed, suspended in a gelatinous protein mixture to allow growth in three dimensions and given a liquid feed. Together, these conditions enabled the cells to develop and grow. Detailed characterisation showed that the resulting structures had self-assembled into “mini-guts” and contained all the cell-types found in the colon in the human body.
- The size of a single “mini-gut” PDO
A “mini-gut” PDO will reach a size of about 80 – 100 microns in diameter if grown for about 5 – 7 days in the lab with appropriate growth conditions. This is equivalent to the width of a human hair and a third of the diameter of a full stop in a printed newspaper, which is about 300 microns. A PDO of this size will contain between 60 – 100 individual cells of the many different types of cells that line the gut. These structures may continue to grow in culture until they are around 10 times that size (1mm) or until they can no longer self-sustain.
- PDOs have been established from organs including the breast, liver, lung, kidney, pancreas and prostate
There are potentially as many types of PDOs as there are different tissues and organs in the body. Each of these types of PDOs will require maintenance conditions reflecting the different function of these cells in the body. They will form different shapes and sizes depending on the tissue of origin.
- PDOs may enable treatment tailored to individuals
When treated with drugs, the effect on cancer PDOs mirrors the patient response. (Vlachogiannis et al., 2018). However, a drug that is effective for one person may not work as well for another due to additional factors.As technology advances, it may one day be possible to produce PDOs from biopsy samples rapidly enough to identify tailored treatments for individual patients i.e., “personalised medicine”. Companies in the US like SEngine (Seattle, WA) and Tempus (Chicago, IL) have already started to do this.Furthermore, using matched normal and diseased PDOs from the same patient may alert us earlier to unwanted, patient-specific, off-target effects.
- PDO shape
PDOs from individual patients and contrasting tissue types can be very dissimilar in appearance. Some PDOs are spheroidal in shape and others have protrusions. Internally, as well, there are differences. Many PDOs have a single cavity or “lumen”, some have several, particularly when the PDO is from cancer tissue where there is abnormal growth of cells.Scientists have observed that the treatment of PDOs with drugs can result in a marked change of morphology which may be related to the effectiveness of the drug. (Badder et al., 2020). Work is progressing to identify what these changes signify and if they can be quantified. This sort of assay could then be used to identify subtle effects that would be missed in live/dead cell counts. The latter, cell-killing assays are commonly used in 2D cell-based platforms. They have however, been shown to be poor predictors of clinical efficacy due to their failure to accurately represent human biology.
- PDOs have the potential to reduce the numbers of animals used in drug discovery
Using fully representative human organoids in compound screening, together with appropriate assays to identify and predict the effect of the treatment in the clinic, will lead to the accurate selection of potential candidates for further development. This will reduce the numbers that need to be tested for toxicity in animals and the likelihood that they will ultimately fail in clinical trials. This may be as much as 97% in cancer drug discovery according to an article by Julia Belluz for Vox in 2019.
- PDOs can be used for multiple research purposes
Experimental studies can be performed on PDOs that aren’t possible on the patients themselves (for example, treatment with drugs and genetic manipulation). They are therefore very useful for research purposes. Amongst other possibilities, PDOs also enable research into the early onset of disease, organ development and drug resistance
- PDOs and “regenerative medicine”
Regenerative medicine refers to the replacement, engineering or regeneration of faulty human cells that are not working as they should. For example, in cystic fibrosis (CF), patients have problems digesting food properly. If PDOs were derived from a patient and the faulty CF genes in the patient’s “mini-guts” corrected, these could then be grown at scale and re-introduced back into the patient, to restore or improve gut function. (E. de Poel et al., 2020).
- Organoid Cell Atlas
This is a world-wide, collaborative effort to examine the cells within PDOs and organoids derived from other sources. This will be part of the Human Cell Atlas whose mission is to create a detailed database relating to the form and function of every cell in the human body and their relationships to each other. This will be the basis for understanding human health and for diagnosing, monitoring and treating disease.Patient Derived Organoids are the research platform of the future. Their clinical relevance will lead to an improved understanding of the biology of the human body. Greater success rates in drug discovery will reduce the cost of drug development and increase the number of effective drugs on the market. Directed treatments will result in less off-target toxicity, reducing the length of hospital stays, improving survival rates and ultimately, the quality of life for patients.
What Cellesce does with PDOs
We offer PDO development and manufacturing services using our patented bioprocess and unique bio-processor technology. Our PDO products in assay ready formats are sold to drug discovery companies and institutions worldwide.
If you would like to talk to us about using PDOs for your research, please feel free to contact us. We look forward to hearing from you.