Bioanalytics Core Leslie M Shaw PPMI Annual Investigators Meeting, - - PowerPoint PPT Presentation

Bioanalytics Core

Leslie M Shaw PPMI Annual Investigators Meeting, NYC May 8, 2013

11/16/2011 1

Bioanalytics Core-2013

• Pilot study manuscript accepted for publication in

JAMA Neurology

• Preparations for analyses of CSF Aβ1-42, t-tau and p-

tau181 in PPMI baseline samples, and baseline, 6 and 12 month longitudinal samples using validated xMAP AlzBio3 immunoassay (all longitudinal samples on same 96 well plate

• Aliquot comparisons; compare current lot of

immunoassay reagents with that used in 2012

11/16/2011 2

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Association of cerebrospinal fluid Ab1-42, t-tau, p-tau181 and alpha-synuclein levels with clinical features of early drug naïve Parkinson’s disease patients; a cross- sectional study By Ju-Hee Kang, David J Irwin, Alice S Chen-Plotkin, Andrew Siderowf, Chelsea Caspell, Christopher S Coffey, Teresa Waligórska, Peggy Taylor, Sarah Pan, Mark Frasier, Kenneth Marek, Karl Kieburtz, Danna Jennings, Tanya Simuni, Caroline M Tanner, Andrew Singleton, Arthur W Toga, Sohini Chowdhury, Brit Mollenhauer, John Q Trojanowski, Leslie M Shaw and the Parkinson’s Progression Marker Initiative

Transfer of PPMI CSFs

(Initial 102 CSF samples for statistical analysis)

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15 Clinical Centers 120 CSF samples 44 Controls 76 Patients 5 F/U CSFs 39 BL HC CSFs 63 BL PD CSFs 4 BL SWEDD CSFs 9 F/U CSFs 102 CSFs for statistical analysis Frozen CSFs

Association of cerebrospinal fluid Aβ1-42, t-tau, p-tau181 and α-synuclein levels with clinical features of early drug naïve Parkinson’s disease patients

J-H Kang, DJ Irwin, AS Chen-Plotkin, A Siderowf, C Caspell, CS Coffey, T Waligórska, P Taylor, S Pan, M Frasier, K Marek, K Kieburtz, D Jennings, T Simuni, CM Tanner, A Singleton, AW Toga, S Chowdhury, B Mollenhauer, JQ Trojanowski, LM Shaw and the Parkinson’s Progression Marker Initiative*

Objective: Evaluate baseline characteristics and relationship to clinical

features of CSF Aβ1-42, t-tau, p-tau181 and α-SYN in PD patients and matched healthy controls (HC) enrolled in PPMI. Methods: CSF biomarkers were measured by xMAP-Luminex platform and ELISA in HC (N=39) and PD (N=63).

Accessioning and statistical analyses of PPMI CSF biomarker dataset

• Simultaneous download of PPMI data for the first 106 study subjects

6/30/2012 by IU and UPenn

• Agreed upon a statistical analysis plan
• Statistical analyses done at UPenn, SAS script sent to IU for replication
• All results in concordance following this process and exchanges of

detailed analyses

• Data assembly, incorporation into draft manuscript
• Manuscript circulated amongst the primary authors and the study site

investigator/authors for suggestions, edits, further discussions of the data

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Statistical Analyses

• Software: SAS ver. 9.3
• Group comparison

— Mann-Whitney U test (2 groups)

— Kruskal-Wallis test with Dunn’s multiple comparison (3 groups)

• Association of CSF biomarker levels and clinical variable

— Multiple linear or logistic regression model (stepwise selection)

with adjustment for confounders (Age, Gender and Education)

• Chi-square test for difference in percentage of subjects
• Pearson correlation

HC (N = 39) PD (N = 63)

P value

Age, years (95% C.I.)

58 ± 13 (54 – 63) 62 ± 10 (59 – 64) 0.2390*

Sex, F/M (% of Male)

18/21 (53.8) 24/39 (61.9) 0.4216#

Education, years (95% C.I.)

16.8 ± 2.4 (16.0 – 17.6) 16.4 ± 2.5 (15.8 – 17.0) 0.1517*

Age at diagnosis, years (95% C.I.)

• 61.1 ± 10.0 (58.6 –

63.7)

• Disease duration, median years

(range)

• 0.4 (0.0 – 2.6)
• Number of subjects with CSF

Hgb > 200 ng/mL

6 18 0.1271#

Demographics of the PPMI subjects

*Mann-Whitney U test; #Chi-square test for HC vs. PD.

*Mann-Whitney U test

¶PR: Right putamen, PL: Left putamen, CR: Right caudate, CL: Left caudate, N=39 for HC, N=62 for PD

Clinical characteristics of the PPMI subjects#

#Data were updated based on PPMI database (06.30.2012)

HC (N = 39) PD (N = 63) p value* H & Y stage 0.03 ± 0.16 1.65 ± 0.51 < 0.0001 UPDRS III motor score 1.6 ± 2.7 22.6 ± 7.6 < 0.0001 Mean tremor score 0.05 ± 0.13 0.53 ± 0.32 < 0.0001 Mean PIGD score 0.01 ± 0.04 0.24 ± 0.26 < 0.0001 UPSIT score 35.1 ± 3.4 21.9 ± 8.1 < 0.0001 Striatal binding ratios (Mean values) PR¶/PL/CR/CL PR/PL/CR/CL <0.0001 1.38/1.39/2.06/2.05 0.62/0.64/1.35/1.34 MoCA (95% C.I.) 28.4 ±1.0 (28.0 –28.7) 27.2 ± 2.0 (26.7 – 27.7) 0.0054 Semantic fluency 53.8 ± 12.1 49.5 ± 10.6 0.0565 WMSIII-LNS$ test score 12.1 ± 2.8 11.0 ± 2.0 0.0510 SDMT 48.6 ± 11.2 41.9 ± 8.9 0.0051 HVLT-R total recall 9.0 ± 1.6 8.2 ± 1.5 0.0077 HVLT-R delayed recall 9.9 ± 2.3 8.3 ± 2.3 0.0004

Comparison of CSF Biomarker levels between HC and PD #

#Data were updated based on PPMI database (06.30.2012)

HC (N = 39) PD (N = 63) P value# Aβ1-42 (pg/mL) 242.8 ± 49.95 (226.7 – 259.0)* 228.7 ± 45.63 (217.2 – 240.2) 0.0466 t-tau (pg/mL) 53.9 ± 19.33 (47.6 – 60.1) 46.1 ± 24.71 (39.8 – 52.3) 0.0276 p-tau181 (pg/mL) 24.9 ± 8.45 (22.2 – 27.6) 21.0 ± 7.83 (19.0 – 23.0) 0.0093 t-tau/Aβ1-42 ratio 0.240 ± 0.141 (0.195 – 0.286) 0.215 ± 0.157 (0.176 – 0.255) 0.0451 p-tau181/Aβ1-42 ratio 0.113 ± 0.075 (0.089 – 0.138) 0.099 ± 0.063 (0.084 – 0.115) 0.1482 p-tau181/t-tau ratio 0.491 ± 0.160 (0.439 – 0.543) 0.543 ± 0.263 (0.477 – 0.609) 0.6820 α-syn (pg/mL) 1264 ± 425.7 (1126 – 1403) 1082 ± 611.1 (928 – 1235) 0.0120

*Mean ± S.D. (95% confidence interval); #Mann-Whitney U test.

MDS-UPDRS Subsection used to classify Tremor or PIDG-dominant phenotype

 Mean tremor score (11 items)

: UPDRS II – 1) Tremor : UPDRS III – 2, 3) Postural tremor (both upper extremities), 4, 5) Kinetic tremor (both upper extremities), 6-10) Resting tremor (4 extremities and lip/jaw), 11) Rest constancy

 Mean Postural Instability & Gait Disturbance (PIGD) score (5 items)

: UPDRS II – 1) Walking and balance, 2) Freezing : UPDRS III – 3) Gait, 4) Freezing of gait, 5) Postural stability

Tremor dominant (TD), or PIGD dominant phenotype

• Ratio of tremor/PIGD score ≥ 1.15: Tremor dominant
• Ratio of tremor/PIGD score ≤ 0.90: PIGD dominant
• 0.90 < Ratio of tremor/PIGD score < 1.15: Intermediate type
• If PIGD score is 0, but tremor score > 0: Tremor dominant

CSF biomarkers according to clinical phenotype in PD patients

12 *PIGD vs. TD; Mann-Whitney U test

# P<0.05 versus HC by Kruskal-Wallis test with Dunn’s multiple comparison

Biomarkers PIGD-PD (N=14) TD-PD (N=43) p value* HC (N =39) IND-PD (N=6) Aβ1-42 (pg/mL) 211.4 ± 45.0# 236.2 ± 46.8 0.0323 242.8 ± 50.0 215.5 ± 25.0 t-tau (pg/mL) 39.3 ± 28.27# 50.3 ± 24.01 0.0527 53.9 ± 19.33 31.2 ± 9.97 p-tau181 (pg/mL) 18.0 ± 6.74# 22.5 ± 8.17 0.0387 24.9 ± 8.45 17.7 ± 4.97 α-syn (pg/mL)a 892.8 ± 542.4# 1185 ± 649.6 0.0587 1264 ± 425.7 782.6 ±150.1 α-syn (pg/mL)b 766.3 ± 446.3# 1122 ± 451.8 0.0286 1267 ± 443.5 775.9 ± 184.8 t-tau/Aβ1-42 ratio 0.211 ± 0.213 0.225 ± 0.145 0.1089 0.240 ± 0.141 0.151 ± 0.072 p-tau/Aβ1-42 ratio 0.093 ± 0.059 0.104 ± 0.068 0.2247 0.113 ± 0.075 0.083 ± 0.026 p-tau/t-tau ratio 0.617 ± 0.398 0.513 ± 0.217 0.7597 0.491 ± 0.160 0.588 ± 0.164

aα-syn was measured in total subjects, or bsubjects with CSF hemoglobin level of < 200 ng/mL

Summary of multivariate regression analyses

• Multivariate regression analysis: lower Aβ1-42 (p=0.0383) and p-tau181 (p=0.0015) are significantly

associated with PD diagnosis, but other biomarkers or their ratios are not.

• For clinical variables in PD, α-syn (p=0.0081) was significantly associated with the MDS-UPDRS III

motor score and t-tau was marginally associated (p=0.0424).

• We found that lower levels of CSF Aβ1-42 and p-tau181 were significantly associated with a higher PIGD

risk.

Correlation between AD biomarkers and α-synuclein:

t-tau p-tau181 Aβ1-42 Total PD HC

Association of cerebrospinal fluid Aβ1-42, t-tau, p-tau181 and α-synuclein levels with clinical features of early drug naïve Parkinson’s disease patients

J-H Kang, DJ Irwin, AS Chen-Plotkin, A Siderowf, C Caspell, CS Coffey, T Waligórska, P Taylor, S Pan, M Frasier, K Marek, K Kieburtz, D Jennings, T Simuni, CM Tanner, A Singleton, AW Toga, S Chowdhury, B Mollenhauer, JQ Trojanowski, LM Shaw and the Parkinson’s Progression Marker Initiative* Results: Significantly lower concentrations of all measured CSF biomarkers and t-tau/ Aβ1-42 ratio were seen in PD compared to HC, lower α-SYN was associated with a higher risk of PD and decreased CSF p-tau181 associated with increased UPDRS motor score. Notably, lower CSF Aβ1-42 was associated with the postural instability-gait disturbance-dominant phenotype which associates with a more rapid cognitive decline and poor prognosis compared to tremor-dominant patients. There is a significant correlation between α-SYN and t-tau & p-tau in PD and HC subjects. Interpretation: We demonstrate that CSF Aβ1-42, t-tau, p-tau181 and α-SYN have value for diagnosis and assessment of disease progression in early-stage PD. Further investigations will test the predictive performance of CSF biomarkers for disease progression.

Bioanalytics Core 2013

• Analyses of the entire PPMI BASELINE CSFs, n=645

and CSFs for those 223 subjects whose 6 month & 12 month CSFs are collected

• AlzBio3 immunoassay for Aβ1-42, t-tau and p-tau181 at

UPenn and α-SYN by ELISA at Covance

• Data analyses, qc, data upload expected by mid to

late summer, 2013.

• Statistical analyses will test hypotheses based on the

pilot study

Acknowledgements: John Trojanowski and the Penn PPMI Bioanalytics Core members Ju Hee Kang, Teresa Waligorska and Sarah Pan, our PPMI study participants, and the amazing PPMI Team!

PPMI Funding Partners

PPMI is sponsored and partially funded by The Michael J. Fox Foundation for Parkinson’s Research. Other funding partners include a consortium of industry players, non-profit organizations and private individuals.

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Scatter plots of CSF biomarker levels in PD

• vs. HC

α-syn Aβ1-42 t-tau p-tau181 Ratios