Purpose: We queried whether comprehensive genomic profiling using a next-generation sequencing– based assay could identify novel and unanticipated targets of therapy for patients with relapsed invasive lobular carcinoma (ILC). Experimental Design: DNA sequencing (Illumina HiSeq 2000) was conducted for 3,320 exons of 182 cancer-related genes and 37 introns of 14 genes frequently rearranged in cancer on indexed, adaptor-ligated, hybridization-captured libraries using DNA isolated from formalin-fixed paraffin-embedded sections from 22 histologically verified ILC. Results:A total of 75 genomic alterations were identified with an average of 3.4 alterations per tumor (range, 1–6), of which 35 were actionable for an average of 1.59 actionable alterations per patient (range, 0–3). Nineteen of 22 (86%) of the ILC samples harbored at least one actionable alteration. Six (27%) cases featured alterations in ERRB2 including 4 (18%) with ERBB2 mutation, 1 (5%) with an ERBB2 gene fusion, and 1 (5%) with an ERBB2 copy number gain (amplification). The enrichment of ERBB2 mutations/fusion in CDH1-mutated ILC (5 of 22, 23%) compared with the 5 ERBB2 mutations in a series of 286 non-CDH1-mutated breast cancers from which the ILC cases were obtained (5 of 286, 2%) was significant (P 1⁄4 0.0006). Conclusions: Comprehensive genomic profiling of relapsed CDH1-mutated ILC revealed actionable genomic alterations in 86% of cases, featured a high incidence of ERBB2 alterations, and can reveal actionable alterations that can inform treatment decisions for patients with ILC. Clin Cancer Res; 19(10); 2668–76. 2013 AACR.

Non-acquired immunodeficiency syndrome (AIDS)-defining neoplasms are being increasingly recognized in patients infected with the human immunodeficiency virus (HIV). The incidence of Hodgkin's disease and seminoma has recently been reported to be increasing in these patients. This article describes the second case of breast cancer in an HIV-infected male patient. A total of 11 cases of coincident breast cancer and HIV infection have previously been reported. It may be prudent to consider breast cancer in the differential diagnosis of an axillary mass in an HIV-infected patient.

CD44 is a family of cell surface proteins implicated in adhesion interactions and tumor metastasis. Multiple CD44 mRNA isoforms arise from alternative splicing of variant exons (termed v1-v10). We recently discovered a novel CD44 mRNA isoform in human papillary thyroid cancers featuring a junction between subsegments of exons 4 and 13 (v8). The sequence ACAG was repeated at both the donor and acceptor sites in the genomic DNA (G. Ermak et al., Cancer Res., 56: 1037-1042, 1996). We used reverse transcription-PCR to characterize expression of this isoform in a panel of thyroid lesions. In addition, we assayed three cryopreserved human breast cancers and two samples of normal breast tissue (from female subjects who had undergone cosmetic mammoplasty) to determine whether a similar isoform is present in breast carcinomas. Levels of the novel isoform were up-regulated in 88% of the goiters, adenomas, and papillary cancers, but were undetectable in cases of thyroiditis and absent or low-level in four samples of normal thyroid tissue. The three breast cancers each yielded a 546-bp PCR product that was not detected in normal breast tissue. The PCR product from one of the breast cancers was cloned, and sequence analysis revealed a novel mRNA isoform featuring a junction between exon 3 and an internal site within exon 13 (v8). The sequence GCTTCAG was repeated at both the donor and acceptor sites in the genomic DNA. These results show that human thyroid and breast tissues contain novel CD44 mRNA isoforms featuring unusual rearrangements at repeated sequences. Further studies are warranted to determine whether the expression of this class of isoforms correlates with growth status.

Mucinous carcinoma (MC) of the breast is characterized by abundant extracellular mucin and by variable epithelial cellularity. Since some MCs are extremely hypocellular, we questioned the validity of biochemical (BIO) assays in these tumors. We analyzed paraffin-embedded tissue from 34 cases of MC of the breast for quantitative estrogen receptor (ER) and progesterone receptor (PR) using immunohistochemistry (IHC) on the Cell Analysis System CAS-200. Of the 34 cases, 31 (91%) were positive for ER, whereas 18 (53%) were positive for PR. In 21 cases the quantitative ER and PR were assayed biochemically by a dextran-coated charcoal method. Using the BIO results as the "true" values, the sensitivity of IHC for ER and PR was 100% and 78%, and the specificity was 13% and 64%, respectively. The low specificity of the values obtained by IHC was attributed to the fact that eight cases were "falsely" false positive (negative by BIO and positive by IHC) for ER and/or PR. Review of the histologic patterns of all 21 cases showed that 7 of the 8 falsely false positive cases were significantly hypocellular (epithelial cellularity 5-20%) as compared to the remaining cases (epithelial cellularity 20-75%). We conclude that immunohistochemical analysis of ER/PR status using image analysis in MC is a sensitive method, with the ability to detect receptor content when their concentration might be too low to be demonstrated by the conventional method as a result of sparse cellularity.

The tumor suppressor protein, p53, protects somatic cells against the accumulation of genomic alterations. Cells harboring mutant or inactivated wild-type p53 protein are at risk for the development of genomic instability. Nuclear accumulation of p53 protein is associated with the stepwise dedifferentiation of papillary carcinoma. We asked whether nuclear p53 accumulation is associated with two known indicators of poor prognosis in papillary carcinoma. We studied 55 consecutive papillary cancers (28 from Russia, and 27 from upstate New York). Nuclear p53 immunoreactivity was assessed using a monoclonal antibody, DO-1, on Formalin-fixed paraffin-embedded specimens. The DNA index was determined by computerized image analysis of Feulgen-stained sections. Nearly all cases were well differentiated and none were associated with distant metastases or extrathyroidal invasion. All primary lesions were less than 4 cm in diameter, and almost all patients were female. Nuclear p53 immunoreactivity was associated with a high-risk group characterized by two known indicators of poor prognosis: age > 50, aneuploid DNA content, or both. In the high-risk group (N = 24) 33% of cases displayed nuclear p53 positivity, compared with only 6% in a low-risk group (N = 31) which lacked both features (P = 0.015, two-tailed Fisher exact test). Nuclear accumulation of immunoreactive p53 protein is associated with two established indicators of poor prognosis in papillary carcinoma of the thyroid. This result is consistent with the idea that aberrations in p53 function are associated with the stepwise loss of differentiation in this cancer.

The first reported cases of Kaposi’s sarcoma (KS) in homosexual men appeared in 1981 and heralded the onset of the AIDS epidemic (1,2). A year later the first four cases of non-Hodgkin lymphoma (NHL) were reported (3). In 1983 the first cases of primary central nervous system lymphoma (PCL) were described (4). By 1985, NHL was added to KS and PCL as index AIDS-defining neoplasms (5). The incidence of PCL and NHL in particular is sharply on the rise (6,7). All three AIDS-defining neoplasms are characterized by higher grade lesions, more advanced stage, and shorter survival when compared to similar tumors in non-HIV infected patients. The epidemiology and clinical characteristics of these neoplasms is well described. (8–10). Today epidemic KS, PCL and NHL represent major causes of morbidity and mortality in AIDS patients.