SUPPLEMENTARY MATERIALS

NURTURE principal investigators (in bold) and site study personnel

Australia: Monique Ryan, M.D., Kate Carroll, DPT, Katy de Valle, Daniella Villano,

Eppie Yiu, M.B.B.S., FRACP, PhD; China: Paul Wuh-Liang Hwu, M.D., PhD, Yin-Hsiu

Chien, M.D., PhD, Jia-Ying Hsu, RN, Hsi-Wen Huang, MS; Yuh-Jyh Jong, M.D.,

D.M.Sci., Tai-Heng Chen, M.D., Po-Ching Chou, M.D., Yun-Hui Chou, Hao-Wei Chung,

M.D., Jong-Hau Hsu, M.D., W-Y Hsu, M.D., Wen-Chen Liang, M.D., Hsiang-Hung Shih,

M.D., Yi-Ching Wu; Germany: Janbernd Kirschner M.D., Sabine Borell, M.D., Matthias

Eckenweiler, M.D., Ronja Haller, Inka Lotz, Astrid Pechmann, M.D., Sibylle Vogt,

Sabine Wider; Italy: Enrico Bertini, M.D., Michela Catteruccia, M.D., Giulia Colia, PT,

Giorgia Copponi, PhD, Adele D’Amico, M.D., PhD, Susanna Livadiotti, M.D., Giorgio

Tasca, M.D., PhD; Valeria Sansone, M.D., Emilio Albamonte, M.D., Alessandra Di Bari,

Luca Mauro, Marta Moscardi, Francesca Salmin; Qatar: Tawfeg Ben-Omran, M.D.,

Nader Al-Dewik, PhD, Mariam Al-Mureikhi, M.D., Fatima Al-Mesaifri, M.D., Patricio

Ayroso, Khalid Ibrahim, M.D., Ahmed Yagoub Magboul, M.D., Sejal Nariani, Ajith Jacob

Ninan, Mahmoud Othman, M.D.; Turkey: Haluk Topaloğlu, M.D., Nesibe Eroğlu, M.D.,

Göknur Haliloğlu, M.D., Ayşe Karaduman, PhD, Gülşah Merve Örsel, Öznur Yılmaz, PT,

PhD; United States: Russell Butterfield, M.D., PhD, Fumiko Alger, Liz Diaz, Melissa

McIntyre, PT, DPT, Nicole Rausch, Amelia Wilson, PT, DPT; Thomas Crawford, M.D.,

Agnes King Rennie, Meghan Moore, PT, DPT, Jessica Nance, M.D., Kerry Vela, PT,

DPT; Darryl De Vivo, M.D., Joshua Cappell, M.D., PhD, Kayla Cornett, PhD, Sally

Dunaway Young, PT, DPT, Jacqueline Montes, PT, EdD, Rachel Salazar, PT, DPT;

Richard Finkel, M.D.; Debbie Berry, RN, Matthew Civitello, MPT, Dawn Cook, RN, Erin

Davis, OTR/L, Craig Johnson, D.O., Wendy Leon, RN, Annie Lim, D.O., Jenna Turner,

PT, Melisa Vega, BS, Fabiola Weber-Guzman, M.D., Jennifer Wilson, RN, Matthias

Zinn, M.D.; Nancy Kuntz, M.D., John Dao, Braeden Dolan, Kristin Krosschell, DPT;

Julie Parsons, M.D., Kaylee Booker, Terri Carry, PT, Lauri Filar, Joanne Janas, M.D.,

Hannah Johnson, Kyle Kusmik, Vi Tran, Michele Yang, M.D.; Kathryn Swoboda, M.D.,

Maria Soledad Hermann, M.D., Sarah Simeone, PT, DPT, Elise Townsend, DPT, PhD,

PCS, Kendall Trautman

Supplementary Table 1

Timing of data collection for assessments.

Study period Screening Treatment

Study dayD -21 to

D -1 D1D15

(±1D)D29

(±1D)

D64, D183, D302, D421, D540, D659, D778 (±7D)

D365, D700 (±7D)

D897 (±7D)

WHO motor milestones x x x x

HINE-2 motor milestonesa x x x

Neurological examinations (includes HINE-1 to Day 778)

x x x x x x x

CHOP INTENDb x x x xUlnar CMAPc x x x xPeroneal CMAPd x x xGrowth parameters x x x xpNF-H x x x x xSafety assessments x x x x x x xAEse x x x x x x xSibling assessmentsf x x

AE, adverse event; CHOP INTEND, Children’s Hospital of Philadelphia Infant Test of Neuromuscular Disorders; CMAP, compound muscle action potential; D, day; HINE-2, Hammersmith Infant Neurologic Examination, Section 2; pNF-H, phosphorylated neurofilament heavy chain; WHO, World Health Organization.a Collected at screening, Day 64, and each study visit through Day 778.b Collected at screening and study visits as shown until a maximum score of 64 was achieved.c Collected at screening, Days 64, 183, 302, 365, 700, then alternate dosing visits.d Collected on Days 64, 183, 302, 365, 700, then alternate dosing visits.e Collected continuously throughout the study.f Collected at screening, on Day 868 or the early termination visit, and on Day 897.

Supplementary Fig. 1. CSF pNF-H levels in NURTURE infants by study visita

CI, confidence interval; CSF, cerebrospinal fluid; pNF-H, phosphorylated neurofilament heavy chain; SMN2, survival motor neuron 2.

pNF-H levels were evaluated using a pNF-H ELLA from ProteinSimple. 7.46 pg/mL was used as the imputed value if the pNF-H concentration was below the limit of quantitation. Baseline pNF-H values in NURTURE infants were obtained on Study Visit Day 1, either prior to nusinersen administration or four hours post-dose. aTime points with n≥5 included. The number of decimal places reported in summary statistics is not indicative of biomarker assay precision or sensitivity.bGeometric mean values at Day 1, 64, and 778 for 2 SMN2 copy children were 20,139.2 pg/mL, 684.6 pg/mL, and 475.2 pg/mL, respectively.cGeometric mean values at Day 1, 64, and 540 for 3 SMN2 copy children were 951.5 pg/mL, 219.5 pg/mL, and 148.3 pg/mL, respectively.

Supplementary Fig. 2. (A) Ulnar and (B) peroneal CMAP amplitude.

A.

B.

CMAP, compound muscle action potential; SE, standard error; SMN2, survival motor neuron 2.Time points with n ≥ 5 included. Normative values for peroneal nerve stimulation of the tibialis anterior muscle are not available.aMiller RG, Kuntz NL. J Child Neurol 1986;1:19–26.

Supplementary Fig. 3. (A) Association between Day 64 weight for age or (B) Day 64 ulnar CMAP amplitude and achievement of WHO walking alone.

A.

B.

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1WHO 99th percentile for age of achievement for development in healthy children: WHO Multicentre Growth Reference Study Group. Acta Paediatr Suppl 2006;450:86‒95.

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Supplementary Fig. 4. (A) Alanine aminotransferase and (B) aspartate aminotransferase levels over time.A

B

ALT, alanine aminotransferase, AST, aspartate aminotransferase; SMN2, survival motor neuron 2.Time points with n≥5 included.

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Supplementary Fig. 5. Creatine kinase levels over time.

SMN2, survival motor neuron 2.

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Supplementary Fig. 6. Creatinine levels over time.

SMN2, survival motor neuron 2.

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Supplementary Fig. 7. Cystatin C levels over time.

SMN2, survival motor neuron 2.

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Supplementary Fig. 8. Platelet levels over time.

SMN2, survival motor neuron 2.

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