1. Fragile X: A Family of Disorders Dianne M. McBrien, MD Clinical Associate Professor of Pediatrics University of Iowa Hospitals and Clinics

3. Sequence of bases codes for proteins Proteins then facilitate or inhibit various reactions related to physiology Change or abnormality in the sequence is called a mutation

4. Triplet repeat expansion mutation Normal number of CGG repeats at site is less than 50 Premutation consists of 55 to 200 repeats The mutation consists of >200 repeats Number of repeats can increase with generations

5. What is fragile X? Family of genetic conditions Mutations in what is now known as FMR1 gene, discovered 1991 Xq27.3 Involves unstable series of trinucleotide repeats

6. FMR1 status can be: Normal (<45 repeats) Grey zone allele (45-54 repeats) Premutation (55-200 repeats) Full mutation (>200 repeats) When we talk about a patient with fragile X syndrome, we are generally referring to an individual with the full mutation. When we talk about a patient who is a carrier, we are referring to an individual with the premutation.

7. What makes the premutation expand to a full mutation in the next generation? Female carrier Number of repeats (more repeats, more unstable) Association with AT base pairs

8. What makes the premutation expand to a full mutation in the next generation? Female carrier Number of repeats (more repeats, more unstable) Association with AT base pairs

9. Full expansion of the gene induces methylation, which silences FMRP production The cognitive and behavioral phenotype is attributed to the complete or partial loss of FMRP

10. What does FMRP do? Binds mRNAs Helps to transport them along the neuron to where they are needed Inhibits translation of these templates until the right signal arrives Helps in synaptic plasticity

11. What does FMRP do?

14. FXS: Problems associated with the full mutation Cognitive deficits Autistic-like features Severe social anxiety AD/HD Behavioral problems Sleep disruption

15. Other medical issues Hypotonia Seizures in at least 17% of males Ear and sinus infections Joint hypermobility Flat feet Mitral valve prolapse PWS-like phenotype in some patients

16. Connective tissue dysplasia cont’d. Hernias Recurrent ear infections Frequent sinusitis Mitral valve prolapse Unusually soft skin

17. Cognitive function Range is quite wide 50-60% within moderate to severe range of MR Function of how methylated (silenced) FMR1 gene is as well as possible mosaicism Characteristic neuropsychological profile

18. Strengths and weaknesses Strong verbal labeling and comprehension—may understand more than rest of abilities would suggest Strong at long-term memory and simultaneous processing Weak at several measures of short-term memory and arithmetic Executive function poor

19. Strengths and weaknesses Strong verbal labeling and comprehension—may understand more than rest of abilities would suggest Strong at long-term memory and simultaneous processing Weak at several measures of short-term memory and arithmetic Executive function poor

20. Autism in FXS More problems with eye contact than children with autism and no FXS dx More social relatedness Extremely high degree of social anxiety

21. Speech and language Poor articulation consistent with cognitive age Rapid, dysrhythmic rate—”cluttering” Self-repetitions (palilalia) c/w autistic individuals without FXS

22. Girls and women with FXS Physical features are milder than in males, more common in patients with full mutation Borderline or mild MR IQ, LD, attentional problems, impulsive behavior, poor eye contact in girls with FM

23. Characteristic appearance May not be present in infants and young children May develop over time Absence of which cannot be used to rule out FXS

24. Premutation carriers Mutation is between 55 and 200 repeats Premutation carriers once thought to be asymptomatic

25. Fragile X tremor-ataxia syndrome (FXTAS) Seen in 25-30% men > 50 years old who have the premutation Limb and truncal ataxia, tremor, cognitive symptoms Misdiagnosed as Parkinson’s disease

26. Premature ovarian insufficiency (POI) 25% of women with premutation Infertility Reduced bone density Irregular menses Menopause prior to 40 years

27. Other problems associated with the premutation ADHD, autism spectrum d/o, learning issues Social anxiety, phobias, and depression SLE, other autoimmune disease Thyroid dysfunction Hypertension Chronic muscle pain syndrome

29. What’s going on? FMRP mRNA in WBCs 2 to 8 times normal High levels of FMRP mRNA in FXTAS inclusions Not toxic in itself Thought to induce a state of oxidative stress  ”heat shock” proteins

30. Targeted treatments: Promise, uncertainty

31. Animal models for fragile X

32. The mGlur5 theory Activation of mGlur5  LTD of synaptic responses LTD much more pronounced in the KO mouse than in wild mouse (Huber et al, 2002) Neuropsychiatric phenotype response to exaggerated mGlur5 response

33. Knockout mouseWild type mouse

34. In the knockout mouse, mGlur5 blockers have shown: correction of dendritic spinal abnormalities correction of seizure threshold improvement in several measures of learning

35. mGlur5 blockade: Human trials Fenobam (Berry- Kravis et al, 2009): Improved communication, eye contact, PPI deficit AFQ056: Response on ABC, CGI, as well as a measure of repetitive behaviors

36. mGlur5 antagonist clinical trials R04917523 (FX trial in adults, now closed) Same compound in children—the Foxtail study, still enrolling

37. Phase II study Double blinded, randomized placebo trial Three arms: 0.5 mg, 1 mg, and placebo WISC-IV, ADOS, Vineland, Social Responsiveness Scale, ADOS, ABC, CGI-S and CGI-I, VAS, Vineland, RBANS, Caregiver Burden Inventory

38. The role of GABA Lower levels of GABA-A receptors Abnormally low GABA activity in amygdala GABA-B agonists block glutamine release Arbaclofen

39. Potent GABA-B agonist Improvements in ABC social withdrawal score, Vineland Play and Leisure Scale, and Visual Analog Scale, as well as improvement on CGI Well tolerated, with minimal side effects Trial was discontinued abruptly

40. Lithium Inhibits GSK3B, which is a kinase Overactive in the KO mouse Li normalizes its levels Reverses a number of behavioral abnormalities in the KO mouse, along with dendritic spinal abnormality and seizure threshold

41. Human trials of lithium in FXS 15 patients with FXS Improvements in Total ABC score, Hyperactivity, Inappropriate Speech and Lethargy (Social Withdrawal) subscales The Maladaptive Behavior subscore from the VABS A parent visual analog scale for target behaviors CGI scale RBANS list learning task Subgroup continued on Li for a year with persistent improvement on ABC-C and VABS ( Berry-Kravis et al. 2008 )

42. Minocycline Treatment of KO hippocampal cell culture improves dendritic spine maturity Treatment of nursing KO dams improves hippocampal dendritic spine anatomy in pups Reduced anxiety in exposed pups

43. Human trials of minocycline Paribello et al: Positive effects in a group of boys 13 years and older Currently a controlled trial going on at UC-Davis for children from 3.5 to 16 years

44. Center for Disabilities and Development University of Iowa Fragile X Clinic