II:3 III:1 III:2 III:3

Transcription

1 Assessment (Range) II:3 III:1 III:2 III:3 Age 54 Age 59 Age 27 Age 32 Age 23 Age 28 Age 18 Age 23 TRS (0-144) NA UPDRS part III (0-108) UHDRS (0-124) SARA (0-40) INAS (0-16) TCS (0-80) Co-registration method Calcification increase (Yes/No) Yes Yes No Yes Table e-1: Motor and radiological scores of the subjects belonging to a kindred with the c.26t>g (p.leu9arg) mutation in the PDGFB gene. Mean time of clinical follow-up was 5.5 years and time between brain CT scans 4.8 years. INAS: Inventory of non-ataxia symptoms; SARA: scale for the rate and assessment and rating of ataxia; TCS: Total calcification score; TRS: Tremor rating scale; UHDRS: Unified Huntington s disease rating scale; UPDRS: Unified Parkinson s disease rating scale.

2 Subject Age at onset of migraine with aura Age at onset of movement disorder Type of movement disorder II: Postural tremor III: Tremor and impaired dexterity* III: Action tremor and postural tremor III: Postural tremor and impaired dexterity Table e-2: All the patients report tremor and reduced dexterity as initial symptoms. Chorea emerged in the course of this study in generation III. * This patient had onset of speech difficulties a year before.

3 Cognitive domain Neuropsychological test Subject III:1 Subject III: (z score) 2015 (z score) 2015 (z score) Brief cognitive status examination MoCA (Montreal cognitive assessment) NA 29 (0.73) 27/30 MMT (Mini mental test) 29/30 28/30 27/30 General intellectual ability IQ Ravens progressive matrices Verbal episodic memory RAVLT (Rey Auditory Verbal Learning Test) 51 (-0,16) 67 (1,61) 42 (-1,29) learning RAVLT retention 15 (1,52) 15 (1,5) 11 (-0,08) Visuo-spatial episodic memory ROCFT (Rey Osterrieth Complex Figure Test) 22 (-0,5) 22 (-0,3) 13 (-3) * immediate recall ROCFT (delayed recall) NA 21,5 (-0,4) 13 (-3) * Working memory Digit span/wais 11 (-1,33) 10 (-1,67)* 10 (-1,67) * Spatial/visual construction ROCFT copy 32 (-0,27) 32 (-0,27) 23 (-3,17) * Block Design/WAIS 56 (1,33) 47 (0,33) 20 (-1,33) Verbal concept formation Similarities/WAIS 25 (0,33) 26 (0,33) 22 (0) Word fluency FAS/COWAT (Controlled Oral Word 33 (-0,7) 56 (1,45) 30 (-0,98) Association Test) Picture naming ability BNT (Boston Naming Test) 49 (-2,46)* 51 (-1,75)* 48 (-2,82) * Information processing speed SDMT (Symbol Digit Modalities Test) 58 (-0,41) 59 (-0,28) 38 (-2,71) * Executive function TMT B (Trail Making Test) 76 (-1,08) 77 (-0,56) 111 (-3,28) * Motor speed FT (Finger-tapping test) dominant hand 40 (-0,74) 50 (0,98) 51 (0,97) FT non-dominant hand 47 (1,14) 48 (1,32) 46 (0,96) Table e-3: Summary of cognitive features in two patients from the F13 family affected by the L9R mutation in the PDGFB gene. A z score -1.5 SD is compatible with a significant deficit (*).

4 Language assesment Test Subject III: Raw score (zscore) 2015 Raw score (zscore) Picture naming task BNT 49 (-2,46) * 51 (-1,75) * COWAT (Verbal fluency) Letters (FAS) 33 (-0.4) 42 (-0.03) Category (Animal names) 31 (2.0) 27 (0.33) Verbs 20 (0.8) 17 (0.40) Repetition task Subtle language disorders (1) 12 (-2.9)* 14 (-2.38)* Word sequence production Months forward (seconds) 6 (-1.3) 5.5 (-1.1) Months backward (seconds) 17 (-0.6) 13.5 (-0.3) Language comprehension Subtle language disorders (1) 18 (-0.7) 18 (-0.7) Table e-4: Language features of the index case (III: ) in the F13 family. *A qualitative analysis of these tests indicated the emergence of paraphasias as a new feature. BNT: Boston naming test. COWAT: Controlled oral word association test. A z score -1.5 SD is compatible with a significant deficit (*). References: 1. Holmbro E, Olsson M. Unpublished monography, Department of speech therapy, Gothenburg University

5 Subject and radiological exam Date of the study Slice thickness used in CT scan (mm) CT scan or MRI manufacturer, magnetic field II:3 First CT June Philips Brilliance 40 Second CT February GEHC Discovery CT750HD First MRI July 2011 GEHC Signa HDxt, 1.5 T Second MRI February 2015 GEHC Discovery 750, 3 T III:1 First CT September Philips Brilliance 40 Second CT September GEHC Discovery CT750HD, First MRI December 2009 Siemens Symphony, 1.5 T Second MRI September 2014 GEHC Discovery 750, 3 T III:2 First CT October GEHC, Lightspeed VCT Second CT August GEHC, Discovery CT750HD First and only MRI August 2015 GEHC, Discovery 750, 3 T III:3 First CT September GEHC, Lightspeed VCT Second CT September GEHC, Discovery CT750HD First and only MRI September 2014 GEHC, Discovery 750, 3 T Table e-5: Dates of radiological investigations, slice thickness and manufacturer. T: Tesla.

6 Measurements of HU in respective ROI ROI size in mm (non-calcified/calcified area) 120 kvp (First CT exam) 60 kev 65 kev 70 kev 75 kev Total change in HU kev 20x /+2 (non-calcified) 8x /+0.8 (non-calcified) 8x /+295 (calcified area) 8x /+34 (calcified area) 10x /+113 (calcified area) Table e- 6: Comparison of Hounsfield units (HU) measurements using different kev levels on second CT of subject III:3. A clear progression is seen in all the calcified areas regardless of kev level. HU: Hounsfield units.

7 Brain region and subject II:3 III:1 III:2 III:3 (max score) first exam last exam first exam last exam first exam last exam first exam last exam TCS (80) Cerebellum (10) Midbrain (10) Thalamus (10) Lentiform nucleus (10) Caudate nucleus (10) White matter (10) Medulla oblongata (5) Pons (5) Vermis (5) Cortex (5) Table e-7: Two patients, III:1 and III:3, were found to have progression of brain calcifications when the total calcification score (TCS) was used. Calcifications in the white matter was very variable: periventricular in II:3, III:2 (punctate appearance) and III:3. In III:1 it was found in the frontal white matter. L: left; R: right.

8 Structure ROIs (N) R/L first exam last exam Difference in percentage (average of ROIs) R/L Difference in HU (average of ROIs) R/L II:3 Cerebellum 3/0 0 0 NA NA Thalamus 1/1 0 0 NA NA Lentiform n 3/3 4/4 4/4 35 (30-44) /25 (6-46) 53 (38-73) /38 (13-61) Caudate n 0/0 0/0 0/0 NA NA White matter 2/3 4/3 4/3 47 (41-52) /33 (29-40) 53 (45-61) /19 (11-25) Cortex NA NA III:1 Cerebellum 2/3 2/2 3/3 44 (41-46) /58 ((-3) (15-20) /25 ((-2)-40 Thalamus 2/2 2/2 3/3 22 (19-24) /24 (22-25) 11 (9-12) /11 (10-11) Lentiform n 2/2 5/5 5/5 29 (2-56) /4 ((-13) (4-137) / 4 ((-21)-35) Caudate n 2/1 5/5 5/5 6 ((-70)-82) /29-37 ((-124)-49 / 13 White matter 2/0 3/1 3/2 14 (12-25) /NA 12 (11-13) /NA Cortex NA NA III:2 Cerebellum 3/1 2/2 2/2 15 ((-4)-44) /4 5 ((-3)-5)/2 Thalamus 2/2 3/3 3/3 15 (8-21)/9 ((-15)-32) 8 (4-13)/3 ((-7)-13 Lentiform n 3/3 5/5 5/5-3 ((-11)-6) / 10 ((-35)-54-6 ((-21)-9) / 11 ((-133)-153)

9 Caudate n 2/2 5/5 5/5 2 ((-2)-6) / 41 ((40-41) 1 ((-1)-5) /32 (31-33) White matter 1/2 2/2 2/2-6/15(14-16) -3/4(4-5) Cortex NA NA III:3 Cerebellum 2/2 3/3 3/3 33 (24-42) /40 (39-40) 20 (17-23) / 29 (27-31) Thalamus 2/2 2/0 2/2 8 (6-10)/13(13-13) 3 (2-4) /5(5-5) Lentiform n 3/ 5/5 5/5 46 (19-70)/34(18-64) 65 (16-98) / 74 (29-123) Caudate n 2/ 5/5 5/5 11 (9-12) / 12 (6-15) 5 (4-7)/ 7(3-9) White matter 4/4 3/3 3/3 16 ((-5)-34)/22(14-31) 40 ((-5)-125) /19(8-35) Cortex NA NA Table e-8: Two patients, III:1 and III:3 were found to have progression of brain calcifications when the total calcification score (TCS) was used. Three patients had measurable progression in density of the calcifications with the co-registration method. The numbers of ROIs were placed different between the anatomical locations and varied from 1 to 4 depending on the size of the calcifications. We did not find evidence of calcifications in the midbrain, vermis, pons or medulla. N: nucleus. L: left; R: right. NA: not applicable due negative HU or related to to small, to faint or lack f calcifications. * Only average values over 10 Hounsfield units (HU) are considered to display increased density.

10 Patient Low values* HU / HA (mg/cm 3 ) High values* HU / HA (mg/cm 3 ) II:3 75/50 650/450 III:1 75/ /850 III:2 75/ /850 III:3 75/ /1000 Table e-9: Dense calcifications with corresponding HU and hydroxyapatite (HA). Calcifications that appear white in a normal brain windowing display a wide spread of densities/concentrations. *Values are approximates from scatterplots.

11 Parameter/metabolite (Reference values) Case II:3 Age 59 Case III:1 Age 32 CSF-albumin NA 198 (Ref age y: <320 mg/l) CSF-albumin/S-albumin (Q alb) NA 4.6 (Ref age y: <7.1) Case III:2 Age * (Ref age10-30 y: <290 mg/l) 7.4* (Ref age y: < 6.5) Case III:3 Age (Ref age y <290 mg/l 6.1 (Ref age y: < 6.5) CSF-IgG (mg/l) NA CSF-IgG index NA (< 0.7) CSF-β-amyloid NA ( > 450 ng/l) CSF-phospho tau NA ( < 60 ng/l) CSF-tau NA (< 400 ng/l) CSF-CXCL13 NA < 1.0 < 1.0 < 1.0 ( < 7.8 ng/l) CSF-NfL NA * ( < 380 ng/l) P-24S-OH-cholesterol ( ng/ml) P-27-OH-cholesterol (Reference ng/ml) CSF-24S-OH-cholesterol NA 0, (< 3 ng/ml) CSF-27-OH-cholesterol NA < (< 1.5 ng/ml) CSF-7α-OH-3-oxo-4-cholestenoic acid (< 22 ng/ml) NA Table e-10: Markers of neurodegeneration, oxysterol levels in plasma (P) and cerebrospinal fluid (CSF) in the F13 family harboring the L9R mutation in the PDGFB gene. * Indicates abnormal value. Patient III:2 had elevated albumin CSF/serum ratio (Q alb) and patient III:3 had elevated NfL level. Oxysterols were normal in the four patients. Analysis performed in December 2009, all the others were performed in NfL = Neurofilament light chain.

12 Mutation in the PDGFB gene (exon number) c.3g>a (signal peptide) Ethnicity (Previous designation)* Unknown a (F10) Nr of symptomatic subjects with calcifications Nr of asymptomatic subjects with (without) calcifications Headache Movement disorders Psych/Beh features Cognitive decline 4 NR Yes At least in one NR At least in one Other features WMA other than calcification s Ref Dyslexia in one N (1) c.26t>g (signal peptide) c.356t>c (exon 4) Swedish (F13) Brazilian (B) 4 0 Migraine with aura Yes in all Recurrent depressions, substance abuse in one In two Language difficulties in index case 3 2 with migraine only Migraine In one case NR NR Language difficulties N (1) and present work N (1) c.433c>t (exon4) c.439c>t (exon 4) c.445c>t (exon 4) Serbian b (S) 6 1 NR Hyperkinesias in 5, parkinsonism in 1 French c 1 0 Migraine Laryngeal dystonia German b (F8) Depression and anxiety in 2 subjects Executive deficits NR N (1) NR NR Dysphagia Mild (2) 5 1 Yes NR NR NR Vertigo N (1) c.726g>c (exon 6) Deletion of exon 3, 4 and 5 c.3657c>t (exon 4) Japanese 2 0 NR Gait disorder Psychosis Mild impairment French b (F) 9 3 NR Hyperkinesias and parkinsonism Psychosis and apathy French c 1 0 NR Mild ataxia Bipolar disease English 3 (1) NR Ataxia and Psychosis posturing and Executive impairment Mild impairment NR N (3) Seizures in 1 Varying degrees depression Table e-11: Summary of the known the known 9 PDGFB mutations, radiological penetrance and phenotypic features. Most mutations occur in exon 4 which make it a hot spot. * Designation in reference 1. a CT-scan on one of the mutation carriers was not performed, clinical data in this family was very limited. b mutations with reduced clinical penetrance. c Sporadic mutations. d These WMA in a 77 years old female suggested vascular origin. e One mutation carrier lacks brain calcifications and was asymptomatic, however her MRI displayed mild WMA. e This subject lacks cortical calcifications. WMA: white matter abnormalities. NR: no reported. (1,4) NR Widespread d (5) NR NR Mild in 1 subject e (6)

13 References: 1. Keller A, Westenberger A, Sobrido MJ, García-Murias M, Domingo A, Sears RL, et al. Mutations in the gene encoding PDGF-B cause brain calcifications in humans and mice. Nat Genet Sep;45(9): Nicolas G, Jacquin A, Thauvin-Robinet C, Rovelet-Lecrux A, Rouaud O, Pottier C, et al. A de novo nonsense PDGFB mutation causing idiopathic basal ganglia calcification with laryngeal dystonia. Eur J Hum Genet EJHG Oct;22(10): Hayashi T, Legati A, Nishikawa T, Coppola G. First Japanese family with primary familial brain calcification due to a mutation in the PDGFB gene: an exome analysis study. Psychiatry Clin Neurosci Feb;69(2): Le Ber I, Marié R-M, Chabot B, Lalevée C, Defer G-L. Neuropsychological and 18FDG-PET studies in a family with idiopathic basal ganglia calcifications. J Neurol Sci Jul 15;258(1-2): Nicolas G, Rovelet-Lecrux A, Pottier C, Martinaud O, Wallon D, Vernier L, et al. PDGFB partial deletion: a new, rare mechanism causing brain calcification with leukoencephalopathy. J Mol Neurosci MN Jun;53(2): Keogh MJ, Pyle A, Daud D, Griffin H, Douroudis K, Eglon G, et al. Clinical heterogeneity of primary familial brain calcification due to a novel mutation in PDGFB. Neurology Apr 28;84(17):