1. Molecular Biology and Medicine
A personal view

2. Molecular biology in medicine
Understanding of normal and pathological processes in the body
Understanding of drug action and development
Genetics…
… and personalised medicine?

3. Molecular biology and medicine
My past research
Career options

4. Investigation of the Neuronal Calcium Sensor Protein Neurocalcin d

5. Calcium signalling Ca2+ RyR Ca2+-CaM SERCA extracellular space LGC K+
VGC
VGC
PKC
Ca2+
buffer
SOC
Ca2+ ATPase
Ca2+
Ca2+
sensor
NOS
RyR
Ca2+-CaM AC
Na+/Ca2+ exchanger
cAMP
CaMK
IP3R
PDE
Ca2+
buffer
PKA ER
SERCA
Golgi
CaMK
mitochondrion
nucleus

6. Calcium-myristoyl switch in recoverin

7. Proteins and the phospholipid bilayer

8. Inserting a gene into a recombinant DNA plasmid

9. Transforming E.coli with a recombinant DNA plasmid

10. Co-localisation of neurocalcin d and clathrin in HeLa cells
overlay
20m

11. Summary and conclusions
neurocalcin d binds actin, tubulin, clathrin HC and GAPDH
this binding is Ca2+-dependent except for tubulin
Ca2+-myristoyl switch operates in neurocalcin d in vivo
Ca2+ induces neurocalcin d translocation to the TGN/perinuclear region
neurocalcin d co-localises with clathrin in the TGN/perinuclear region at elevated Ca2+
this suggests one role of neurocalcin d may be in intracellular trafficking

12. Why should anyone care??
Certain neuronal calcium sensors have been implicated in aetiology of Alzheimer's disease (AD)
Genome-wide association studies have since shown association of genetic variants of two of these proteins with AD+psychosis and late-onset AD
One of these, VILIP-1, is being tested as a cerebrospinal fluid biomarker and a prognostic marker for cognitive decline in AD
Braunwell K.H., Front Mol Neurosci. 2012; 5: 20.
Kester M.I. et al, 2015, Alzheimer's Research & Therapy20157:59

13. Splicing Factors in
Retinitis Pigmentosa

14. Retinitis pigmentosa (RP)
progressive retinal degeneration
incidence ~1 in 3500
begins in the periphery, results in narrowing of visual fields, night blindness, severe visual impairment, and blindness in ~1/3 of cases

15. Healthy and RP-affected eye

16. Proteins implicated in RP
Phototransduction cascade components 6
Vitamin A transport/storage 6
Cell adhesion molecules 3
Intracellular transport 5
Transcription factors 3
Structural proteins 2
Splicing factors
Unknown

17. Pre-mRNA splicing

18. Ratio of spliced and unspliced RPGR RNA in control and splicing-factor mutant cell lines
PRP31
PRP8
RPGR – autosomal dominant RP
High level of alternative splicing in ex-in 15
Large number of disease causing mutations in this region

19. Summary and conclusions
Splicing of RPGR is significantly reduced in PRP31 mutant cell lines

20. Obvious why we should care…
"…an important role for pre-mRNA splicing in retinal homeostasis and the pathogenesis of retinal degenerative diseases. The development of novel therapeutic strategies to modulate aberrant splicing, including small molecule based therapies, has the potential to lead to the development of new treatments for retinal degenerative diseases."
E.g. gene augmentation therapy (adding DNA containing a functional version of a mutated gene back into the cell, which produces sufficient amounts of a functioning protein to replace the defective/missing one): work started on PRP31.
Liu M.M. and Zack D.J., Clin Genet Aug; 84(2): 142–149.
Hafler et al, Semin Ophthalmol. 2016;31(1-2):49-52.
Beltran et al, Cold Spring Harb Perspect Med Feb; 5(2).

21. Plasmalogen Function(s) in the Retina

22. Plasmalogens What is the function of plasmalogens in the retina? EPA
DHA AA
What is the function of plasmalogens in the retina?

23. How can we investigate this?
DHAPAT -/- mice (DHAPAT catalyses 1st step in plasmalogen biosynthesis)
reduce/abolish DHAPAT activity in retinoblastoma cells by RNA interference

24. RNA interference
Y79
deficiency in DHAPAT/ADHAPS activity

25. Summary and conclusions

26. Omega-3 fatty acids
omega-3#section3
Anxiety & depression
Eye health
Brain health
Cardiovascular health
ADHD
Autoimmune diseases
Cancer
Sleep
Skin

27. Antioxidants

28. But Dr Go Ogle says it's a brain tumour.

29. Molecular biology in medicine
Understanding of normal and pathological processes in the body
Understanding of drug action and development
Genetics…
… and personalised medicine?
Career options
Clinical academic: "combine working as a specialist doctor with research and teaching responsibilities" (

30. Career options

31. Career options

32. Be a doctor in your heart, and a scientist in your brain