SUPPLEMENTARY INFORMATION
Germ-line and somatic DICER1 mutations in pineoblastoma
Leanne de Kock1,2, Nelly Sabbaghian2, Harriet Druker3, Evan Weber4, Nancy Hamel4,5, Suzanne
Miller6, Catherine S. Choong7, Nicholas Gottardo8, Ursula R. Kees9, Surya P. Rednam10,
Liselotte P. van Hest11, Marjolijn C. Jongmans12, Shalini Jhangiani13, James R. Lupski13, 14,
Margaret Zacharin15, Dorothée Bouron-Dal Soglio16, Annie Huang17, John R. Priest18, Arie
Perry19, Sabine Mueller20, Steffen Albrecht21, David Malkin22, Richard G. Grundy6 and William
D. Foulkes1,2,4,5
Correspondence: Dr. William D. Foulkes at the Department of Medical Genetics, The Lady Davis
Institute, Segal Cancer Centre, Jewish General Hospital, 3755 Cote St. Catherine Road, Montreal,
QC, Canada, H3T 1E2. Email: william.foulkes@mcgill.ca.
SUPPLEMENTARY FIGURES
Supplementary Figure S1:
Supplementary Figure S1 Legend:
Flow chart summarizing the mode of ascertainment of cases, sample acquisition, molecular analysis
and the results of the study. In the section 2 (sample acquisition), boxes shaded in grey indicate
samples that were sequenced by us as a part of the study. Molecular screening of samples contained in
white boxes was performed at other institutions, including at the referring institution (n = 2), Ambry
Genetics (Aliso Viejo, CA, USA) (n = 1), Prevention Genetics (Marshfield, WI, USA) (n = 2), or at
Baylor-Hopkins Center for Mendelian Genomics (Houston, TX, USA) (n = 1)
Supplementary Figure S2:
Supplementary Figure S2 Legend: Loss extends to a large region of 14q in tumours showing loss
of heterozygosity.
Analysis of short tandem repeat (STR) markers for each tumour included one STR marker near
DICER1 and one marker closer to the centromere of chromosome 14 q. A) Genotype analysis of STR
markers visualized using gel electrophoresis of radiolabeled PCR amplicons. The two alleles are
referenced for each marker using black (allele 1) and red (allele 2) bars. In tumours where there is
LOH of one allele compared to the normal cells, the missing allele is always allele 2 (red). N =
normal; T = tumour. B) Graphical representation of the position of the STR markers analyzed and
DICER1 on chromosome 14q. Marker positions and LOH status for each patient analyzed are
displayed to the right of the graphic for each marker. No = no LOH; NI = not informative
(homozygous in normal tissue); LOH = loss of heterozygosity.
SUPPLEMENTARY TABLES
Supplementary Table S1: The comparison of one somatic missense mutation in DICER1 RNase IIIb
domain versus no somatic missense mutation in DICER1 RNase IIIb domain occurring in PinB and
other DICER1-associated tumour types.
Supplementary Table S1: 2x2 Contingency Table - The occurrence of RNase IIIb missense
mutations in PinB vs RNase IIIb missense mutations in other DICER1-associated tumours
Somatic missense mutation in
DICER1 RNase IIIb domain
No somatic missense mutation
in DICER1 RNase IIIb domain
PinB
0
6
Other DICER1-associated
tumours
59
1
Other DICER1-associated tumours included were: Pituitary blastoma (PitB), pleuropulmonary
blastoma (PPB), cystic nephroma (CN), Sertoli-Leydig cell tumours (SLCT) and Wilms tumour (WT)
Fishers Exact Test Results: The two-tailed P value is 7.7x10-8 indicating that the absence of
missense RNase IIIb mutations in PinB is unlikely to be a chance finding. Cases included in the
calculations were those found to possess two DICER1 “hits.” The six PinBs which were incorporated
in the calculations included the one previously reported case [1], one case from the International PPB
Registry (IPPBR) described by Doros et al. [2], case 10, case 11 and case 19 from the current study
which harboured a germ-line DICER1 mutation and LOH of the DICER1 locus, and case 12 which
carried two truncating DICER1 mutations. LOH has been identified in two cases of pituitary blastoma
[3] but only the one case which harboured both a germ-line DICER1 mutation and LOH was included
in the calculation.
References:
PinB
1. Sabbaghian N, Hamel N, Srivastava A, Albrecht S, Priest JR, Foulkes WD (2012) Germline
DICER1 mutation and associated loss of heterozygosity in a pineoblastoma. J Med Genet 49
(7):417-419. doi:10.1136/jmedgenet-2012-100898
2. Doros L, Schultz KA, Stewart DR, Bauer AJ, Williams G, Rossi CT, Carr A, Yang J, Dehner LP,
Messinger Y, Hill DA (1993) DICER1-Related Disorders. In: Pagon RA, Adam MP,
Ardinger HH et al. (eds) GeneReviews. University of Washington, Seattle, Seattle WA,
PitB:
3. de Kock L, Sabbaghian N, Plourde F, Srivastava A, Weber E, Bouron-Dal Soglio D, Hamel N,
Choi JH, Park SH, Deal CL, Kelsey MM, Dishop MK, Esbenshade A, Kuttesch JF, Jacques
TS, Perry A, Leichter H, Maeder P, Brundler MA, Warner J, Neal J, Zacharin M, Korbonits
M, Cole T, Traunecker H, McLean TW, Rotondo F, Lepage P, Albrecht S, Horvath E, Kovacs
K, Priest JR, Foulkes WD (2014) Pituitary blastoma: a pathognomonic feature of germ-line
DICER1 mutations. Acta Neuropathol. doi:10.1007/s00401-014-1285-z
Sahakitrungruang T, Srichomthong C, Pornkunwilai S, Amornfa J, Shuangshoti S, Kulawonganunchai
S, Suphapeetiporn K, Shotelersuk V (2014) Germline and Somatic DICER1 Mutations in a
Pituitary Blastoma causing Infantile Onset Cushing Disease. The Journal of Clinical
Endocrinology & Metabolism 0 (0):jc.2014-1016. doi:doi:10.1210/jc.2014-1016
PPB:
Foulkes unpublished data.
Murray MJ, Bailey S, Raby KL, Saini HK, de Kock L, Burke GA, Foulkes WD, Enright AJ, Coleman
N, Tischkowitz M (2014) Serum levels of mature microRNAs in DICER1-mutated
pleuropulmonary blastoma. Oncogenesis 3:e87. doi:10.1038/oncsis.2014.1
Seki M, Yoshida K, Shiraishi Y, Shimamura T, Sato Y, Nishimura R, Okuno Y, Chiba K, Tanaka H,
Kato K, Kato M, Hanada R, Nomura Y, Park MJ, Ishida T, Oka A, Igarashi T, Miyano S,
Hayashi Y, Ogawa S, Takita J (2014) Biallelic DICER1 mutations in sporadic
pleuropulmonary blastoma. Cancer Res. doi:10.1158/0008-5472.CAN-13-2470
Pugh TJ, Yu W, Yang J, Field AL, Ambrogio L, Carter SL, Cibulskis K, Giannikopoulos P, Kiezun
A, Kim J, McKenna A, Nickerson E, Getz G, Hoffher S, Messinger YH, Dehner LP, Roberts
CW, Rodriguez-Galindo C, Williams GM, Rossi CT, Meyerson M, Hill DA (2014) Exome
sequencing of pleuropulmonary blastoma reveals frequent biallelic loss of TP53 and two hits
in DICER1 resulting in retention of 5p-derived miRNA hairpin loop sequences. Oncogene 0.
doi:10.1038/onc.2014.150
CN:
Foulkes unpublished data
Doros LA, Rossi CT, Yang J, Field A, Williams GM, Messinger Y, Cajaiba MM, Perlman EJ, K AS,
Cathro HP, Legallo RD, Lafortune KA, Chikwava KR, Faria P, Geller JI, Dome JS, Mullen
EA, Gratias EJ, Dehner LP, Hill DA (2014) DICER1 mutations in childhood cystic nephroma
and its relationship to DICER1-renal sarcoma. Mod Pathol. doi:10.1038/modpathol.2013.242
SLCT:
Heravi-Moussavi A, Anglesio MS, Cheng SW, Senz J, Yang W, Prentice L, Fejes AP, Chow C, Tone
A, Kalloger SE, Hamel N, Roth A, Ha G, Wan AN, Maines-Bandiera S, Salamanca C, Pasini
B, Clarke BA, Lee AF, Lee CH, Zhao C, Young RH, Aparicio SA, Sorensen PH, Woo MM,
Boyd N, Jones SJ, Hirst M, Marra MA, Gilks B, Shah SP, Foulkes WD, Morin GB,
Huntsman DG (2012) Recurrent somatic DICER1 mutations in nonepithelial ovarian cancers.
N Engl J Med 366 (3):234-242. doi:10.1056/NEJMoa1102903
WT:
Wu M, Sabbaghian N, Xu B, Addidou-Kalucki S, Bernard C, Zou D, Reeve A, Eccles M, Cole C,
Choong C, Charles A, Tan T, Iglesias D, Goodyer P, Foulkes W (2013) Biallelic DICER1
mutations occur in Wilms tumours. J Pathol. doi:10.1002/path.4196