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Figure 1. Morphological data. (A) The epithelial component of well-differentiated fetal adenocarcinoma consists of cells
harboring clear, glycogen-rich cytoplasmic vacuoles. A sparse fibrous stroma can be noted in the background (hematoxylin
and eosin stain, magnification 40). (B) The epithelium displays mostly membranous, but also multifocal, cytoplasmic and
nuclear immunohistochemical positivity for b-catenin (magnification 600). Images taken before laser capture microdis-
section of the epithelial component (C) and mesenchymal component (hematoxylin and eosin stain) (D). Images taken of the
same areas subsequent to laser capture microdissection of the epithelial component (E) and mesenchymal component
(Arcturus HistoGene stain [Arcturus Bioscience, Inc., Mountain View, CA], magnification approximately 20) (F). Black di-
amonds indicate dissected areas.
Figure 2. Sequencing of genomic DNA obtained after laser capture microdissection of the epithelial and mesenchymal
components of the well-differentiated fetal adenocarcinoma. Mutation locations are indicated by an asterisk, and the wild-
type nucleotide sequence is provided below each chromatogram. (A) The germline DICER1 mutation (c.3540C>A) is present
in both the malignant epithelium (panel I) and background benign mesenchyme (panel II). (B) The somatic DICER1 mutation
(c.5127T>A) is present only within the epithelium (panel I). (C) The somatic b-catenin gene (CTNNB1) mutation (c.98C>G) is
similarly present within the epithelium (panel I). There is a slight suggestion of the c.98C>G mutation in the mesenchymal
component (panel II); the most likely explanation for this is contamination by some interpolated malignant epithelial cells.
March 2016 DICER1 Mutations in Fetal Lung Adenocarcinoma e33
The family demonstrated a typical inherited germ- epithelium is novel. Our results raise intriguing ques-
line DICER1 mutation in blood lymphocyte genomic tions about the relationships between three rare pul-
DNA: c.3540C>A (p.Tyr1180*) (Fig. 2A).2 Sanger monary tumors: PPB, which is an exclusively
sequencing of RNase III domains in tumor genomic DNA mesenchymal malignancy; WDFA, which is an adeno-
from the proband’s three tumors revealed an additional carcinoma, and PB, which is a biphasic carcinosarcoma.
somatic DICER1 mutation in each: WDFA, c.5127T>A PB and WDFA may share CTNNB1 mutations, and our
(p.Asp1709Glu); SLCT, c.5438A>C (p.Glu1813Ala); and results show that PPB and WDFA may share DICER1
MNG, c.5126A>G (p.Asp1709Gly). These are typical mutations. Other molecular events underpinning the
DICER1-associated somatic RNase IIIb “hotspot” muta- malignant epithelial component of WDFA should be
tions affecting critical metal ion binding.5 Laser micro- investigated. Moreover, the potential role of DICER1 in
dissection of malignant epithelium and background PB deserves exploration.
benign stroma (Figs. 1C–1F) in the WDFA detected the
somatic DICER1 mutation exclusively in the epithelium Acknowledgments
(Fig. 2B). The authors thank Dr. Morag Park, Dr. Nicholas Bertos,
Also exclusively in WDFA epithelial tissue, we and laboratory members for assistance with the laser
identified a CTNNB1 mutation, c.98C>G (p.Ser33Cys) capture microdissection and subsequent DNA extrac-
(Fig. 2C). When subjected to immunohistochemical tions and Talia Boshari for help with sample acquisition
staining, the tumor exhibited multifocal aberrant and associated administrative work.
epithelial nuclear/cytoplasmic expression of b-catenin
(see Fig. 1B). There is a slight suggestion of this mutation References
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in a patient with an ovarian Sertoli-Leydig tumor, well-
Discussion differentiated fetal adenocarcinoma of the lung, and fa-
Before our earlier report of this case,2 WDFA had milial multinodular goiter. Eur J Med Genet. 2014;57:
not been reported within the DICER1 syndrome (OMIM 621–625.
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