1. ASCTs could prove to be a useful therapeutic target for neuropsychological disorders Jill Farnsworth GradCon 2016

2. Neurological Disorders Diseases of the brain and spine including nerves that connect them Include: – Alzheimer’s Disease – Parkinson’s Disease – Epilepsy – Stroke – Schizophrenia Many therapeutic targets http://nutri.com/blog/2013/09/neurological-disorders-and-vitamin-d/

3. NMDA Receptors and Disease NMDAR activity = molecular and cellular basis for learning and memory NMDAR hypofunction implicated in schizophrenia – Hyperfunction in stroke Current treatments show little success We propose a novel therapeutic target https://www.nia.nih.gov/alzheimers/scientific-images

4. NMDAR Modulation by ASCT ASCT = Alanine Serine Cysteine Transporters D-serine = NMDAR synaptic coagonist – Needed for NMDAR activation D-Ser is specific to NMDAR – Decreased likelihood of off-target affects – D vs L isoforms D-Ser modulation has therapeutic potential

5. Do ASCTs transport D-serine? Methodology: – Inject hASCT1/2 RNA into oocyte Incubate expressing oocytes in [ 3 H]D-Ser Measure [ 3 H]D-Ser accumulation in oocytes http://www.labtimes.org/labtimes/method/methods/2009_01.lasso

6. Do ASCTs transport D-serine? ASCT1/2 transport D-Ser with micromolar affinity – Frog oocytes expressing hASCT1/2 show radio- labeled [ 3 H]D-Ser uptake Extracellular [D-Ser] present in micromolar quantities – Indicates ASCT as a mediator of [D-Ser] ASCT2 K m = 196+/- 52µM ASCT1 K m = 217+/- 52µM

7. Regulating ASCT Activity Inhibiting ASCT may enhance D- Ser availability – Treat schizophrenia Compound (BiPro) developed here at UM BiPro = (2S,3R, 4R)-4-(biphenyl-4-ylmethoxy)-3-hydroxy-pyrrolidine-2-carboxylic acid)

8. Regulating ASCT Activity BiPro inhibits ASCT1/2 with nanomolar potency – Measured by decrease in D-Ser transport Shows competitive inhibition Specific to ASCT ASCT2 IC 50 = 121+/- 22nM ASCT1 IC 50 = 224+/- 70nM

9. Regulating ASCT Activity BiPro inhibits ASCT1/2 with nanomolar potency – Measured by decrease in D-Ser transport Shows competitive inhibition Specific to ASCT ASCT1

10. BiPro is Physioloigcally Relevant NMDAR activation modulates learning and memory Behavioral phenomenon measured by LTP – Long-Term Potentiation 100 and 300nM BiPro facilitate LTP – Experiments in rat brain visual cortex https://mcauliffeneur493/home/synaptic-plasticity

11. BiPro is Physioloigcally Relevant NMDAR activation modulates learning and memory Behavioral phenomenon measured by LTP – Long-Term Potentiation 100 and 300nM BiPro facilitate LTP – Experiments in rat brain visual cortex

12. Future Directions BiPro’s modulation of [D-Ser] in other cortical areas – Does it facilitate LTP in all areas with NMDARs? Can BiPro rescue NMDAR hypofunction? – Schizophrenia models BiPro and cancer? https://www.nia.nih.gov/alzheimers/scientific-images