RESEARCH - BMHA LOTTERY GRANT- SUMMARY

Lottery Funding – After two application attempts £150,000 was awarded to the BMHA to fund research into the further development of a DNA blood test for MH families, the ultimate aim of such research being to replace the present MH test method which requires an invasive muscle biopsy. The funding began April 2005. The grant employs a full time research technician and a part time administrative assistant.
Advantages – DNA testing if widely used would i) reduce costs to the NHS for MH screening avoiding operating theatre and staff time required for muscle biopsy testing ii) minimise the inconvenience and unease to patients iii) increase numbers of people tested and allow testing of young children and the elderly.
Progress – DNA extracted from blood samples from BMHA members is currently being analysed for mutations in the RYR1 gene, the major MH susceptibility gene. Mutations are being detected using a new analysis process. Independent of the lottery funding, the BMHA have contributed to the purchase of a new machine to allow this analysis to be performed more efficiently.

DETAILS

The BMHA, together with the Leeds MH Investigation Unit, worked for many months to formulate a research project entitled “The advancement of Malignant Hyperthermia (MH) testing by DNA analysis”. This was submitted to the Lottery Research Grant programme in 2002, but unfortunately was turned down.

St James's University Hospital
The BMHA Management Committee decided to have another attempt at obtaining lottery funds for this project. In late 2003, after many more months of work, we were approved. The grant awarded was ~£150,000. We were warned that it would take at least 6 months before we would be able to begin the project, but in the end, for various reasons beyond our control, this turned out to be over a year!

The following are the main areas of work as submitted in our bid.

What is the overall aim of the research project?

Anyone suspected of having had an MH reaction needs to be tested at the Leeds MH Investigation Unit for a definitive diagnosis. Until recently, this involved a muscle biopsy. Guidelines for DNA testing were published in 2001. An initial study by the Leeds MH Unit indicated that DNA diagnosis according to these guidelines is possible for approximately 25% of families. This leaves 75% of families who would not benefit.

The overall aim of the project is to be able to offer blood tests to all families not just the 25% who can benefit at present. This will involve contacting families, collecting blood samples for analysis, searching for new mutations causative of MH and informing families of the results.

Why is there a need for this research project?

An MH crisis results in a great deal of trauma to the patient and their families and considerable expense to the NHS via specialist drugs and intensive care treatment. The original crisis then needs confirmation. The only current method of testing is by a surgical procedure (muscle biopsy), followed by family screening to identify other family members at risk. The Leeds MH Unit is the only testing centre in this country and can only test 6 patients a week (approx. 230 per year). Many families find the journey to Leeds and the idea of an invasive test too much to undertake and it can take months/years for the whole family to be tested. Children cannot be tested by biopsy until they are ~10 years old and elderly people often do not want to travel.
BMHA members have for many years strongly supported research for a simpler blood test, and have raised many thousands of pounds over the last 10 years for this research.

Why the research is important?

MH can be fatal if undetected or treated incorrectly. As it is an inherited genetic problem even families where some members have already been tested, will have other relatives and future generations who will need to receive a definitive diagnosis. The Leeds MH Unit has over 650 MH families registered involving around 5000 individuals whose relatives and descendants would benefit from DNA testing as far fewer members would require a muscle biopsy. Patients are more agreeable to testing via DNA diagnosis, as no surgical procedure is necessary; it only requires a blood specimen and it does not entail travel to the MH Unit. A family’s susceptibility status is more likely to be complete as more family members can be expected to be tested. In contacting as many families as we can through this project, the database can be updated resulting in more effective patient support and better dissemination of information.

Outline of the research

The initial phase will be for Leeds to check through current BMHA members to identify families for whom they do not have a suitable blood sample and these people will be asked to participate. We will then move on to lapsed members and then finally any other UK MH family not known to BMHA. At this stage, to be covered by NHS confidentiality rules, we will appoint an administrator to work within the Leeds Unit alongside a Genetic Technician who would be appointed for the whole 3 year project. 

Progress to date

Year two progress

At the end of year two 153 independent MH families have ongoing genetic investigations being conducted. The patient rebleeding program is largely complete. Response to rebleed request letters sent to BMHA members was not as high as expected. However, the Leeds MH unit has many patient samples already stored which still require genetic analysis to be performed so these were latterly included in the study. Nevertheless, 57% of families included were affiliated with the BMHA through current or previous membership.

Progress with the laboratory work has been good with analysis of 68/153 patient samples almost complete for targeted screening of the RYR1 gene which is associated with MH in the majority of cases. Approximately one third of the analysis is presently complete for the remaining 83 patients being studied. Work has been conducted using a combination of MELT and sequence analysis methods. So far mutations have been detected in 33 families. In 7 families a mutation which may be used in genetic diagnosis of MH has been detected – the Leeds MH Unit is now in the process of contacting these families who will immediately benefit from these findings. In the remaining families 13 mutations previously reported in the literature have been detected and a further 10 novel mutations have been identified.

These findings were recently reported by the research group at the XXVI Annual European MH Meeting held in Siena, Italy at the end of May.

Unfortunately, late last year, the technician ceased working on the project due to an unrelated incident.  The MH Unit seconded another member of staff part time (Chris) and the University provided a technician (Nikla) so that the momentum built up would be affected as little as possible.  It is a great testament to the commitment of those involved with this project that the above work has been achieved despite the difficulties. 

We have also just heard that Rachel will be leaving the unit as she has secured a high-level promotion within the Yorkshire Regional DNA laboratory.  A proposal is currently with the Big Lottery Fund to delay the start of year 3 and we will keep you updated when we have more news. 

 

Year one progress

One of the tests originally proposed for searching for mutations in the patient DNA samples was been superseded by a new MELT analysis process. This works on the basis that when a stretch of DNA is heated, a fragment with a mutation will ‘melt’ at a different rate compared to the same fragment without the mutation. If such a difference is observed the DNA requires further analysis to properly characterise the mutation. This is good method of analysis and will provide the researchers with new work for publication. However, it meant that an additional machine was needed by the laboratory for DNA amplification (PCR machine). This was been funded mainly by BMHA, independent of this project.

At the end of year one of the project, progress met expected targets. The MELT analysis process had been successfully introduced into the laboratory after much careful optimisation. A database was been developed to record and co-ordinate all results for the project. The first group of BMHA members donatde a blood samples for analysis. The first batch of patient DNA samples was analysed for area’s of the RYR1 gene, which from previous investigations appear to be hot-spots for MH related mutations. Some mutations were detected and further characterised.

Further updates will appear when available.

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