Gene, stem and cellular therapies awards

Awarded to William Mason for £21,957 as part of the new projects call 2023.

One of the main functions of the kidney is to filter the blood of impurities, while conserving the important molecules needed for life. Between 1 and 3 children in every 100,000 born worldwide will have a genetic problem in the way their kidney’s filter the blood. These patients do not respond to current therapies, meaning at present, there are no treatments to slow disease progression. For most of these infants, blood filtration becomes so severely impaired that they reach end-stage kidney disease, becoming completely reliant on the high-burden and costly option of dialysis while awaiting a possible kidney transplant. Therefore, alternative therapies that slow disease or support kidney filtration offer a significant way to increase the quality for life of these children.

The site of filtration in the kidney is called the glomerulus, which contains a barrier to aid in filtration. This barrier is made up of blood vessels and two layers of specialised kidney cells that together form a tight filtration mesh, much like a coffee filter, retaining all the important coffee, whilst letting the water and small particles through. Damage to any part of this barrier can result in leaky filtration, which if left untreated can cause kidney failure.

This project will use a new method to grow all three parts of the filtration barrier in a dish, something not possible with traditional techniques, recreating the human glomerulus. This is termed the glomerulus-on-a-chip! We will generate this model in the laboratory and use it to determine what goes wrong in childhood glomerular disease. Then we will use a treatment which has shown promise in pre-clinical animal trials to assess its therapeutic benefit in humans.

Overall, this project will establish a human platform which will allow us to test new therapies for childhood glomerular disease, accelerating alterative therapy discovery.

Awarded to Asma Naseem for £18,000 as part of the new projects call 2023.

Krabbe disease is a rare, serious genetic disease of the nervous system. A genetic disease is caused by changes in an individual's DNA sequence. It is a demyelinating disease in which the protective layering of the nerves called Myelin sheath gets damaged; due to which the nerves cannot function properly.

People with Krabbe disease can't repair their myelin due to changes in a gene that codes for an enzyme required for the maintenance of myelin (known as GALC or galactosylceramidase). Enzymes are proteins that are required for the normal functioning of the human body.

When this protective layer of myelin is damaged, nerve impulses slow or even stop, causing neurological problems which are the major symptoms of Krabbe disease. Infants with Krabbe disease may not show symptoms at birth but soon develop feeding difficulties, muscle weakness, and developmental delays which quickly progress to total loss of motor function, vision, and hearing.

Like many other demyelinating disorders, there is no cure for Krabbe disease, but it can be treated with transplantation when performed immediately after diagnosis (best within 6 months after birth). In this process, blood cells from a healthy donor containing normal levels of GALC enzyme are given to the affected patients to stop the progression of the disease; however, it cannot repair the myelin that has already been damaged.

In this project, we plan to develop a way of correcting Krabbe disease before birth, using a specific DNA-modifying protein packed into a harmless, "empty" virus shell known as a virus-like particle. This virus shell can still get into human cells like a normal virus would but does not cause an infection, and after entering it falls apart to release the cargo (the DNA modifying protein) into the cell which will correct the mutation to allow normal functioning of the gene.

Awarded to Soraia Caetano-Silva for £24,948 as part of the new projects call 2023.

Rare but devastating immune system disorders such as X-linked lymphoproliferative disease (XLP) have limited treatment options. Currently bone marrow transplant is the only curative therapy. For these procedures, blood stem cells (haematopoeitic stem cells) from a healthy donor are infused into the patient where they engraft in the bone marrow space and develop a healthy immune system. However, transplant can be complicated by rejection of the cells or graft versus host disease (GvHD), especially if the donor is not well matched to the patient.

Much needed novel, safer and effective alternative therapies, like gene therapy are evolving. In these a corrective copy of the defective gene is delivered into the patients’ own stem cells or the gene is 'edited' in the patients cells. Gene editing could offer a safer option in some diseases where continuous expression could lead to unwanted effects like an increased risk of cancer. We consider XLP a disease where a gene editing approach to correction may be superior.

Instead of adding a new gene to the patients genome (DNA), gene editing corrects the genes at its own site in the genome. This protocol uses the regulatory signals already available in the surrounding DNA to replicate normal expression.

Here we plan to test the efficacy and compare different ways of delivering these corrective gene templates to blood stem cells focusing on non-integrating lentiviral vectors. We will also compare this system to the current more standard AAV vectors to bring safer and potentially curative gene edited stem cell therapy closer to patients in need. Although this project will focus on developing this platform for XLP, many other severe inherited diseases could benefit from this study in the future.

Awarded to John Anderson for £39,741 as part of the new projects call 2023.

We wish to run an immunotherapy clinical trial for children with brain cancer, which involves gene modification of the patient’s own blood killer cells. Brain cancer is one of the largest clinical unmet needs in paediatric oncology. Each year over 400 children in the UK are diagnosed with a tumour of the brain or spinal cord an of these about 100 children each year receive a diagnosis that will ultimately prove fatal.

Our approach to a new treatment that is more targeted and effective is based on genetically modifying the killer cells of the immune system to redirect them specifically to seek out and kill cancer cells. These types of therapies have been very successful at treating previously incurable blood cancers, but solid tumours like brain cancer have proved much more difficult to treat. One of the reasons that such genetically modified killer cells struggle with solid tumours is that the genetic modification by itself is sufficient to create a state of inactivity termed “T cell exhaustion” which limits their activity before even having a chance to see the tumour. We have designed an approach to get around this by allowing the immune cells to rest and recover from this state of exhaustion. This is achieved by switching off the genetic modification by adding a drug during the manufacturing process.

The herein project is focused on optimising the details of how we make the immunotherapy to ensure that it is rested but highly functional. Parameters we will look at include: (i) scheduling of how and when we add the drug during manufacture, (ii) switch to a more efficient gene-engineering method to enhance expression of the genetic modification, (iii) evaluating different compositions of the conditions in which the genetically modified killer cells are grown.

The output of the project will be an optimised gene-engineered immune cell manufacturing process that we can take forward into clinical trial for evaluation of safety and efficacy in the treatment of childhood brain cancer.

Awarded to Laura Donovan for £40,000 as part of the new projects call 2023.

Central nervous system tumours are the leading cause of cancer related deaths in children. Medulloblastoma (MB) is a type of brain cancer which begins in the lower back part of the brain, called the cerebellum. Although MB can occur at any age, it is the most common, aggressive brain tumour in young children. Four different subgroups of the disease exist, each with differing demographics and clinical outcomes (WNT, SHH, Group3 and Group4). Group3 subgroup shows the highest rate of universally fatal recurrence than other MB subgroups. Therapies at recurrence do not exist, the only option is palliative care. Recurrent MB represents one of the biggest challenges in the brain tumour community and the complete lack of therapeutic options emphasises the need for urgent novel therapies.

Chimeric antigen receptor T-cell (CART) therapy is a very complex and specialised immunotherapy; cells from a patient’s blood, known as T-cells, are altered to recognize and target a specific protein on the surface of cancer cells, effectively killing the cancer without harming normal tissue. CART therapy has shown success in blood cancers, such as leukaemia, but less success in solid tumours, resulting in fatal tumour recurrences. The belief is that the CARTs lack the perseverance with solid tumours. Previous research has demonstrated that CARTs perseverance is affected by genes being turned on and off, a mechanism known as epigenetics. Epigenetic mechanisms are reversible and can be controlled with appropriate drugs, many of which are currently in the clinic, such as Azacytidine.

The lead applicant has demonstrated extraordinary results when CART therapy is used in combination with the epigenetic modifier, Azacytidine. The CART and drug combination effectively treated, and in many cases, cured mice harbouring patient-derived brain tumours, including medulloblastoma. To enable us to move this combined therapy into clinical trials for brain tumours, the mechanism of action between Azacytidine and CARTs must be explored. To evaluate this mechanism, Azacytidine will be used in combination with CARTs in human models of group3 MB and characterised using state-of-the-art sequencing techniques. Results from this project are critical for direct translation into novel, preclinical, immune-competent mouse models of group3 MBs, for the next generation of clinical trials.

Awarded to Giorgia Santilli for £21,644 as part of the new projects call 2023.

X-Linked Agammaglobulinaemia (XLA) is a genetic disease caused by mutations in the genetic code. The genetic code is like a language with instructions that cells use to make up proteins. Any change in a letter or in a sequence of letters can cause defects in protein production, often leading to genetic diseases.

XLA patients, for example, are missing an important protein that is key to the formation of a specific type of blood cells called "B lymphocytes”. These B lymphocytes produce antibodies, tiny molecules that combat viruses and bacteria. Due to the absence of antibodies, individuals with XLA experience frequent infections and often require prolonged hospital stays. The recent COVID-19 pandemic has introduced a new challenge in the treatment of XLA patients, as they are prone to chronic and recurrent Covid infections.

Currently, the only available therapy for XLA is lifelong infusions of antibodies. However, this treatment has limited efficacy and places a burden on the NHS, as it costs approximately £2 million per patient.

B lymphocytes originate from specialised "blood stem cells" that normally reside in the bone marrow. Today, it is possible to isolate these blood stem cells and manipulate them in a laboratory setting to correct the genetic defect. The corrected cells can then be reintroduced to the patient through a process similar to blood transfusion. Once infused, these cells "home" to the bone marrow and commence the production of normal blood cells throughout the patient's lifetime.

Our proposal aims to correct the blood stem cells of XLA patients using a technique called "base editing," which allows scientists to modify individual letters of the genetic code. With BRC funds, we will trial the base editing procedure on blood stem cells from one XLA patient to determine if they can regenerate B lymphocytes and produce antibodies like healthy cells once the genetic defect is corrected. If successful, this approach holds the potential to be a future cure for XLA patients and others with similar genetic disorders.

Awarded to Thomas Whittaker for £23,400 as part of the new projects call 2023.

Genes are the instructions in our DNA that the human body uses to build and repair itself. Genetic diseases happen when a patient has a gene which has an error in it, meaning that when it is read the correct product isn’t made. They often cause serious health problems but can potentially be permanently cured by inserting a replacement gene into the patient’s DNA. One way of doing this involves physically inserting a replacement piece of DNA with the fixed gene on it into the patient’s cells and prompting the patient’s cells to incorporate it into their own DNA. However, human cells have mechanisms that sense if DNA is present in the cell in an unusual way and shut down the cell. Cells have these defence mechanisms to protect against viruses and to recognise if they are damaged, but they can also be activated by gene therapies, meaning that the corrected cells cannot always be successfully transplanted back into the patient.

We want to develop a new way of introducing DNA into the cell that does not cause the cell to shut down. The way that the sensing pathways work for detecting DNA is already well-understood and it should be possible to avoid them by a combination of changes to the DNA itself (these changes are naturally found in human cells and are not dangerous) and improvements in the way the DNA is purified. We have developed a manufacturing process that allows us to produce this DNA at a scale sufficient for further experiments. We are requesting funding to test this modified DNA in human cells – specifically, the stem cells that make the immune system that are given in a bone marrow transplant - to see whether it can allow us to correct them without setting off their DNA-sensing mechanisms.

Project: Liver-directed lentiviral gene therapy for progressive familial intrahepatic cholestasis type 2 (PFIC2).

Awarded to Ziyu Jiang for £96,669.

Project: Rescuing B cell development in patients affected by X-linked agammaglobulinemia.

Awarded to Stefano Gritti for £96,669.

Also part of the Career Development Academy.