Clinical and genetic analysis of recurrent adult-type granulosa cell tumor of the ovary: Persistent preservation of heterozygous <i>c</i>.<i>402C>G FOXL2</i> mutation

<div><p>Background</p><p>Adult-type granulosa cell tumors of the ovary (aGCTs) are rare tumors that represent 2–5% of ovarian malignancies. The prognosis of this tumor is favorable, and it is characterized by slow progression. 10–30% of these tumors recur after 4–7 years of the primary surgery and the 5-year survival rate from the first recurrence is 55%, for the incompletely resected patients. At this time, complete resection is the only prognostic factor for better outcome, and establishing a novel strategy for identification and/or treatment of recurrent tumors is crucial. After the discovery of heterozygous <i>c</i>.<i>402C>G FOXL2</i> mutations in 97% of cases of aGCT, much effort has been made to find the role of the mutation on the pathogenesis of aGCT, however, little is known about the role of the mutation in disease progression.</p><p>Methods</p><p>We analyzed the clinical data of 56 aGCT patients to find a marker of recurrence. In particular, we compared the <i>FOXL2</i> status in 5 matched primary and recurrent samples by immunohistochemistry, and TaqMan allelic discrimination assay to address the role of <i>FOXL2</i> in potential mechanisms of recurrence.</p><p>Results</p><p>The clinical data analysis was consistent with complete resection as an indicator of disease eradication, though the sample size was limited. The genetic analysis showed all the samples, including recurrent tumor samples up to 14 years after the primary surgery, expressed heterozygous <i>c</i>.<i>402C>G FOXL2</i> mutation and the FOXL2 protein expression.</p><p>Conclusion</p><p>This report describes the preservation of heterozygous <i>c</i>.<i>402C>G FOXL2</i> mutation in recurrent aGCTs. This finding adds further credence to the concept that the <i>c</i>.<i>402C>G FOXL2</i> mutation is oncogenic and integral to this disease.</p></div

Histological finding and p53 expression of Pt #5.

<p>(a) The primary tumor of the Pt #5 revealed the bland tumor cells with variably nuclear grooves and a predominant diffuse pattern with minor trabecular and insular pattern which compatible with aGCTs. The mitotic activity was inconspicuous. (b) The recurrent tumor of the Pt #5 showed the similar histological findings as the primary tumor, though the mitotic activity was slightly higher compared to the primary tumor. (c) Primary and recurrent samples from patient #5 showed the moderate to strong p53 staining in the nucleus.</p

TaqMan based allelic discrimination assay.

<p>All samples gave fluorescent signal from both WT and mutant probes consistent with heterozygous c.402 C>G FOXL2 mutation. Each sample was duplicated and the black dot represents the negative control (H2O).</p

Clinical findings of patients with recurrent disease.

<p>Clinical findings of patients with recurrent disease.</p

Risk factors for recurrence (multivariate analysis).

<p>Risk factors for recurrence (multivariate analysis).</p

Patient characteristics.

<p>Patient characteristics.</p

Risk factors for recurrence (Univariate analysis).

<p>Risk factors for recurrence (Univariate analysis).</p

Single-Patient Molecular Testing with NanoString nCounter Data Using a Reference-Based Strategy for Batch Effect Correction

<div><p>A major weakness in many high-throughput genomic studies is the lack of consideration of a clinical environment where one patient at a time must be evaluated. We examined generalizable and platform-specific sources of variation from NanoString gene expression data on both ovarian cancer and Hodgkin lymphoma patients. A reference-based strategy, applicable to single-patient molecular testing is proposed for batch effect correction. The proposed protocol improved performance in an established Hodgkin lymphoma classifier, reducing batch-to-batch misclassification while retaining accuracy and precision. We suggest this strategy may facilitate development of NanoString and similar molecular assays by accelerating prospective validation and clinical uptake of relevant diagnostics.</p></div

Genome-wide copy number profiles of bona-fide ovarian CCC cell lines.

<p>A large range of copy number changes are seen including typical Chr8 gains and Chr17 gains surrounding the CCC biomarker <i>HNF1B</i> gene, see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0072162#pone-0072162-t003" target="_blank">Table 3</a>.</p

PVCA of the OC clinical samples after adjusting batch effect using different methods.

<p>We consider the PVCA plot of the OC clinical samples run in different batches after adjusting BE with different methods. In each plot, percentages represent the variability explained by each factor and first order interaction between factors.</p