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Abstracts

Brazil. The use of the array CGH (aCGH) has aided significantly the genetic diagnosis in recent years. In our laboratory
aCGH combined with conventional karyotyping allowed an
improvement of 20% to 30% in the diagnostic rate of patients
with intellectual disabillity, dysmorphisms, autism and developmental delay. We used the Affymetrix chip Cytoscan HD and
750 K Cytoscan arrays (Santa Clara, CA) and analysis through
the respective software system and for conventional karyotype
(G Banding). We selected 2 cases to report: Case 1: Two
siblings with distinct developmental disturbance phenotypes.
Sibling one is a 7-year-old female with severe developmental
delay phenotype, microcephaly and consanguineous parents.
She presented an interstitial deletion of 12,1 Mb on chromosome 5, arr [hg19] 5p14.3- p15.31 (6,801,589-18,992,827) 1,
featuring the Cri-du-chat syndrome. Her brother, a 12-year-old
male with mild intellectual disability (ID) a duplication of the
exact same size and region that was deleted in the sister was
found arr [hg19] 5p14.3-p15.31 (6,801,589-18,992,827) 3.
Parental aCGH was normal. However, conventional karyotype
of the father revealed 2 reciprocal translocation between
chromosomes 1 and 2, 5 and 7: (46, XY, t(1, 2) (q44;w p23pter) t(5; 7)(p14.3-p15.31; p22). Case 2: A 16-year-old female
with consanguineous parents and normal conventional karyotyping. She presents significant developmental delay, dyslalia, severe ID, autistic traits, irritability, hyperactivity and
autoagression, without obvious dysmorphias and with normal:
MRI, electroencephalogram and CT scan results. Microarray
analysis revealed a homozygous microdeletion of 197 Kbp on
chromosome 8:arr [hg19] 8p22 (15,451,748-15,649,733) 1,
spanning almost the entire TUSC3 gene, except promoter and
first exon (601385*).This is the seventh family with a reported
TUSC3 mutation with mostly non syndromic ID and the second
with the same deletion (Khan et al, 2011). Three of the families
reported in the literature carried a deletion which encompassed
only the TUSC3 promoter region and the first exon, 2 families
presented distinct point mutations and 1 an intragenic duplication. All 7 families presented homozygous mutations, only 1
was not consanguineous however they were from the same
village.
Conclusion: These cases illustrate the importance of integrating conventional karyotyping techniques and microarray in
genetic diagnosis. Both cases were referred for genetic counselling and family research.

Unusual ROS1 Translocation Pattern in a 61 Year-old


Woman with Metastatic Adenocarcinoma of Lung
Hui Chen a, Rajyalakshmi Luthra b, Neda Kalhor a,
John Heymach c, Ronald Abraham b, Meenakshi Mehrotra b,
Bal Mukund Mishra b, Keyur P. Patel b, Rajesh R. Singh b,
Xinyan Lu b
a
Department of Pathology, MD Anderson Cancer Center, Houston,
TX, USA; bDepartment of Hematopathology, MD Anderson Cancer
Center, Houston, TX, USA; cDepartment of Thoracic/Head &Neck
Medical Oncology, MD Anderson Cancer Center, Houston, TX, USA

FISH assay has enabled us to detect gene amplification,


deletion and translocation in solid tumors. However, FISH is
limited to focused genes of interest. Currently the alternative
microarray based genome-wide analysis for copy number aberration and allelic imbalances of oncogenes remains undercharacterized. In this study, we explored genomic copy number

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analysis on a 61-year-old woman with metastatic adenocarcinoma of lung and refractory to chemo- and adjuvant therapies.
Primary tumor from fine needle aspiration of lung and metastatic adenocarcinoma from core biopsy of lymph node were
used for immunohistochemistry (IHC), FISH assay and next
generation sequencing analysis to detect overexpression/
amplification, translocation, and somatic mutations. Additionally, genomic DNA from metastatic tumor was subjected to
molecular inversion probe array (MIP) by OncoScan FFPE
Assay kit (Affymetrix). Genomic copy number and allelic
imbalance calls were performed by OncoScan Console and
data reviewed by OncoScan Nexus Express (BioDiscovery).
Both primary and metastatic tumors were pan negative for
EGFR, KRAS and BRAF mutations and ALK translocation.
FISH analysis demonstrated ROS1 translocation with unusual
double splitting signals suggestive of allelic imbalance in lymph
node metastasis but not in primary tumor. MIP study on lymph
node metastasis showed loss of heterozygosity involving 4
chromosomes/arms including chromosome 6 where ROS1
located. In addition, MIP array detected copy number gains
involving 9 chromosomes/arms. Genome instability with
numerous copy number gain and allelic imbalances, and ROS1
translocation might be one of contributing factors to patient
poor clinical outcome.

A Novel Mutation in Calreticulin (CALR) was Identified


in a Patient of African American Origin with
Thrombocytosis
D.P. Dash a, Sherine Joseph Thomas b
a
Blood Center of Wisconsin, Milwaukee, WI, USA; b Georgia Cancer
Specialists and Northside Hospital Cancer Institute, Atlanta, GA,
USA

Recent studies show that Calreticulin (CALR) somatic mutations provide a diagnostic marker in JAK2/MPL wild type
essential thrombocythemia (ET) and primary myelofibrosis
(PMF) with a mutation frequency of 67% to 71% and 56% to
88%, respectively (Tefferi A and Pardanani A, Nat Rev Clin
Oncol, 2014). In general, CALR mutations are not seen in
Polycythemia Vera (PV), post-PV myelofibrosis or JAK2 V617F
or MPL mutated ET or PMF. Studies showed that CALR mutations may also provide prognostic information and therefore is a
promising molecular marker for diagnosis and prognosis for
patients with myeloproliferative neoplasm (MPN). The exact
mechanism by which CALR mutations produce the myeloproliferative disease phenotype is unknown, but multiple studies have
shown that CALR mutations disrupt the C-terminal endoplasmic
reticulum -retention sequence (KDEL), generate a novel C-terminus, and activate the STAT5 pathway. A 40-year-old female
with persistent moderate thrombocytosis was referred for evaluation. Workup was negative for pseudo-thrombocytosis. She
was noted to have mild iron deficiency. Reactive thrombocytosis
and essential thrombocytosis were considered in the differential
diagnosis. She was referred for clinical testing of CALR mutation
analysis after she was found negative for JAK2. Mutation
screening of CALR Exon 09 by PCR based Sanger sequencing
revealed a novel mutation c.1191_1199del (p.E398_D400del) in
this patient with African American ethnicity. Mutation status in
CALR will aid the diagnosis of ET and PMF patients and risk
stratification.