Estrogen Receptor (ER) and Progesterone Receptor (PR) Immunohistochemistry is Sensitive and Specific for Differentiating Retroperitoneal Leiomyosarcomas With Symplastic-like Features From Their Uterine Mimics.

Background: Patients with HER2+ breast cancer who have residual disease after neoadjuvant anti-HER2 plus chemotherapy have a high risk of recurrence and benefit from adjuvant trastuzumab emtansine (T-DM1). We hypothesize that endocrine-responsive residual tumors after neoadjuvant treatments may have good outcomes among patients receiving only adjuvant endocrine therapy plus trastuzumab. Using paired pre- and post-treatment samples from CALGB 40601 and other neoadjuvant cohorts that did not include adjuvant T-DM1, we investigated survival by pretreatment and residual disease ESR1 and PgR gene expression in CALGB 40601, and by estrogen receptor (ER) and progesterone receptor (PR) immunohistochemistry (IHC) in the other cohorts. We considered endocrine-responsive tumors those with ER and/or PR expression by gene expression or IHC. Methods: CALGB 40601 is a randomized neoadjuvant trial of single vs. dual HER2-targeting (trastuzumab and/or lapatinib added to paclitaxel). We only included those patients who had not suffered from disease progression or death during their preoperative treatments. In total, 77 patients with paired pretreatment and residual disease tumors were profiled by mRNA sequencing and studied. Cutoffs for ESR1 and PgR mRNA expression mimicking clinical positivity were obtained using 265 pretreatment CALGB 40601 tumors (and 1,045 TCGA samples for ESR1). We also examined ER and PR IHC in paired tumors from 202 patients treated at 4 different collaborating institutions; all had residual disease after neoadjuvant HER2 targeting plus chemotherapy. We considered ER- or PR-positive as ≥10% positively staining cells. The primary endpoint was EFS, defined as the time from randomization to event in CALGB 40601 and from the first systemic therapy to event in the 4-institution validation cohort. Results: In 77 patients from CALGB 40601 with paired (pretreatment/residual disease) specimens, 38 (49.3%) had ESR1+/ESR1+ tumors. The EFS was superior in the ESR1+/ESR1+ (n=38) group than in the remaining others (the log-rank test, p=0.011). The 5- and 7-year EFS rates for the ESR1+/ESR1+ (n=38) were 92.1% and 89.2%, whereas the rates were uniformly < 70% in the others. In particular, the 5-year EFS rate among 11 patients with ESR1+/ESR1- tumors was 61.4%. This remained significant in multivariable analysis with clinical stage and treatment arm; the hazard ratio (HR) for EFS in ESR1+/ESR1+ versus all others was 0.29 (95% CI, 0.09-0.90). In ESR1+/ESR1+ tumors, 5-year EFS rates were high for those whose residual disease and also being PgR+ (n=32) or PAM50 LumA or Normal-like (n=34) (93.8% and 97.1%, respectively). In the institutional validation cohort, pretreatment /residual disease ER(+)/ER(+) tumors (n=113) had superior 3-year EFS versus all others (p=0.010). At a median follow-up of 35.9 months, the 3-year EFS rates for ER(+)/ER(+) and all other groups were 96.6% and 82.6%, respectively. This remained strongly significant in multivariable analysis with clinical stage; the HR for EFS in ER(+)/ER(+) versus all other groups was 0.28 (95% CI, 0.09-0.88). Among 46 who also had PR+ in the residual disease, the 3-year EFS was 100.0%. Conclusions: HER2+ patients with ER+ pretreatment and ER+ residual disease after neoadjuvant chemotherapy + HER2-targeting have a very good survival outcome despite not receiving additional anti-HER2 targeting with T-DM1. This may provide a simple mechanism to better tailor therapy within residual disease patients using serial ER measurements. Citation Format: Sung Gwe Ahn, Aranzazu Fernandez-Martinez, Mei-Yin C. Polley, Soong June Bae, Seho Park, Núria Chic, Fara Brasó-Maristany, Benedetta Conte, Valentina Guarneri, Maria Vittoria Dieci, Gaia Griguolo, PierFranco Conte, Joon Jeong, Aleix Prat, Lisa A. Carey, Charles M Perou. A good prognosis of endocrine-dependent tumors among residual invasive cancer after anti-HER2 therapy: CALGB 40601 (Alliance) and validation studies [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS10-02.

Effect of EDTA decalcification on estrogen receptor and progesterone receptor immunohistochemistry and HER2/neu fluorescence in situ hybridization in breast carcinoma

Vascular leiomyoma of the nasal cavity

The distinction of metastatic breast adenocarcinoma (MBA) to the liver from hepatocellular carcinoma (HCC), cholangiocarcinoma (CC), and metastatic adenocarcinoma from other sites may require the use of immunohistochemistry. The use of antibodies directed against estrogen receptor (ER) and progesterone receptor (PR) has been suggested to help make this distinction. To examine the utility of ER and PR immunohistochemistry in the distinction of MBA from HCC, CC, and other metastatic adenocarcinomas in the liver. Ninety-two previously characterized hepatic neoplasms were identified, including HCC (n = 14), CC (n = 16), and metastatic tumors from breast (n = 17), colorectal (n = 14), pancreatic (n = 15), and esophageal/gastric (n = 16) origins. For all cases of metastatic tumor, the primary tumor was reviewed to verify the diagnosis. All tumors were graded as well, moderately, or poorly differentiated. Estrogen receptor and PR immunohistochemical staining was performed on all cases and evaluated by 2 pathologists. Immunoreactivity for ER was identified only in MBA, with 6 (35%) of 17 cases positive. Positive immunoreactivity for PR was not restricted to MBA, but was seen in HCC, CC, esophageal/gastric, and pancreatic metastases. Positive immunostaining with PR was nearly as frequent in poorly differentiated carcinomas of nonbreast origin (3/16 cases, 19%) as in poorly differentiated breast carcinomas (2/8 cases, 25%). Progesterone receptor exhibited poor specificity and sensitivity for the distinction of MBA from HCC, CC, and other metastatic adenocarcinomas. Estrogen receptor exhibited poor sensitivity for MBA, although the specificity was good. The finding that PR positivity was present with a similar frequency in poorly differentiated tumors of breast and nonbreast origin limits the usefulness of this marker. Therefore, ER and PR staining have limited utility in the distinction of MBA from HCC, CC, and other metastatic adenocarcinomas in the liver.

Prognostic Significance of Progesterone Receptor Immunohistochemistry in Endometrial Carcinoma

Digital quantification of Ki-67 and PHH3 in the classification of uterine smooth muscle tumors

Benign uterine leiomyomata (LMA) are hormonally responsive neoplasms. To the authors' knowledge, little is known regarding the hormonal expression patterns of leiomyosarcomas (LMSs) of the uterus. The objective of the current study was to assess the immunohistochemical (IHC) patterns of estrogen receptor (ER), progesterone receptor (PR), and androgen receptor (AR) expression in LMA and LMS of the uterus using a tissue microarray. The authors also sought to assess the prognostic value of ER, PR, and AR expression in LMS. Between January 1991 and March 2001, 25 patients were identified with primary uterine LMS for whom tissue was available. A tissue microarray was created with 3 representative cores from each of the LMS cases, as well as from 19 cases with benign uterine LMA. IHC staining was scored as follows: negative (0-1) and positive (2-3). Outcome analyses were performed for LMS only. First recurrence was determined from the time of the initial diagnosis. Survival was determined from the time of the initial diagnosis to last follow-up. ER, PR, and AR positivity in LMA compared with LMS was as follows: ER, 78% versus 40% (P = 0.03); PR, 88% versus 38% (P = 0.001); and AR, 32% versus 40% (P = 0.75). There was no difference noted with regard to IHC expression based on stage of LMS. The median overall survival for patients with LMS was 23.5 months (95% confidence interval, 19.5, NR). When adjusted for stage, PR and AR were found to be predictive of a lower risk of recurrence (P = 0.01 and P = 0.035, respectively). ER, PR, and AR were not found to be associated with overall survival after adjustment for stage. Tumor stage was found to be associated with survival (P = 0.005). The rate of ER and PR expression was found to be significantly less in uterine LMS compared with LMA. ER, PR, and AR expression was observed in approximately 30-40% of uterine LMS cases. In the current series, PR and AR expression appeared to be associated with disease-free survival but were not found to correlate with overall survival.

BACKGROUND: Recently objective quantification of immunohistochemical estrogenreceptor (ER) and progesterone receptor (PgR) staining in breast cancer by image cytometry has been predominantly performed by measuring the area of positivelystained cells. However, in sample preparations of immunostained hormone receptors, both the stained area and the intensity of staining vary. In this study, we performed quantification of the stained area by measuring, tailing intensity using image cytometry. METHODS: Quantitative analysis of ER and PgR immunohistochemistry was performed using image cytometry. The obtained values were presented as % of positive staining (% PS). Comparison of % PS with values obtained by EIA and with clinicopathological features was performed. RESULTS: The % PS values and the natural logarithm of the EIA levels of the hormone receptors showed a significant positive correlation for both ER and PgR. The concordance of the results obtained by the two methods was 96.3% for ER and 73.7% for PgR. The ER-% PS values of postmenopausal patients were significantly higher than those of premenopausal patients, whereas the PgR-% PS values of the former group were significantly lower than those of the latter group. CONCLUSION: The quantification of ER and PgR in immunostained preparations using % PS as a parameter was reproducible and showed a high correlation with values obtained by EIA. It was shown that only menopausal status affects hormone receptor levels when analyzing the relationship between % PS measurements and clinicopathological features.

The authors have noted anecdotal cases of extrauterine leiomyosarcomas (LMS) with estrogen receptor (ER) and progester-one receptor (PR) immunoreactivity. However, there are few studies that have compared ER and PR immunoexpression in LMS of uterine and extrauterine origin. The authors obtained a representative formalin-fixed, paraffin-embedded tissue block from cases of uterine LMS (n = 15) and extrauterine LMS (n = 16) from the archives of the Cleveland Clinic Foundation and performed immunohistochemical staining for ER and PR. Staining was evaluated by 2 observers in a semiquantitative manner using the following scale: 0, no nuclear staining; 1+, 1 to 25% of nuclei stained; 2+, 26 to 50% of nuclei stained; 3+, 51 to 75% of nuclei stained; 4+, 76 to 100% of nuclei stained. The majority of uterine LMS stained for ER (13 of 15, 87%), PR (12 of 15, 80%), or both ER and PR (12 of 15, 80%), with most cases showing 3+ or 4+ positive staining. For the extrauterine LMS cases, staining for ER was seen in 4 of 16 cases (25%), staining for PR was observed in 2 of 16 cases (13%), and staining for both ER and PR was seen in 2 of 16 cases (13%). One extrauterine LMS showed 4+ coexpression of ER and PR, but the remaining extrauterine cases showed only 1+ ER and/or PR immunoreactivity. These data suggest that most uterine LMS coexpress ER and PR, and most extrauterine LMS do not stain for these antigens. However, a subset of extrauterine LMS are ER and/or PR immunoreactive. This raises the possibility that hormonal manipulation may be beneficial in the treatment of these therapeutically recalcitrant tumors.

Steroid receptors in ovarian carcinoma: Immunohistochemical determination may lead to new aspects

Steroid receptors in ovarian carcinoma: Immunohistochemical determination may lead to new aspects

Estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor (HER2) concordance between immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH), and Oncotype DX, a commercially available RT-PCR-based assay which recently began reporting biomarker results was assessed. ER concordance was 98.9% (262/265), Pearson correlation coefficient (r) = 0.42, and Spearman's rank correlation (ρ) = 0.25. Positive percent agreement for ER was 98.9% (262/265). One patient with discordant ER results was not offered hormone therapy based on the preferential use of Oncotype DX. PR was concordant in 91.3% (242/265), r = 0.80, ρ = 0.75, and Cohen's kappa (κ) = 0.63. Positive percent agreement for PR was 90.5% (218/241) and negative percent agreement was 100% (24/24). HER2 concordance was 99.2% (245/247), r = 0.35, ρ = 0.28, and κ = 0.12. Positive percent agreement for HER2 was 0% (0/2) and negative percent agreement was 100% (245/245). Of the three FISH HER2-amplified cases, two were negative and one was equivocal, and all FISH HER2-equivocal cases (n = 3) were negative by Oncotype DX. Patients that were FISH HER2-amplified, Oncotype DX HER2-negative did not receive trastuzumab. Although our results demonstrated high concordance between IHC and Oncotype DX for ER and PR, our data showed poor positive percent agreement for HER2. Compared to FISH, Oncotype DX does not identify HER2-positive breast carcinomas. The preferential use of Oncotype DX biomarker results over IHC and FISH is discouraged.

To assess the immunohistochemical profile of the atypical nuclei in leiomyoma with bizarre nuclei and compare with benign and malignant counterparts. 26 cases of uterine smooth muscle tumors including 12 leiomyosarcoma(LMS), 10 leiomyoma with bizarre nuclei (LBN) and 4 smooth muscle tumor with uncertain malignant potential (STUMP) were selected using whole tissue sections for this study and analysis. Six cases of ordinary leiomyoma were included as benign control group. All representative section were stained for P53, Ki67, estrogen receptor and progestrone receptor. Analysis was carried out using SPSS 16.0 for windows software. Six out of 12 cases of LMS showed strong and diffuse nuclear staining with p53 antibody (50%). In contrast none of STUMPs and only one case of LBN cases showed focal positive reaction with P53. Percentage of positive cells for ki67 in LMS was 14.92 while only 0.85% of cells in LBNs was labeled with Ki67 proliferative marker. (P<0.001). Regarding steroid hormone receptors a significant loosing trend was found in these receptors from benign toward malignant tumors through LBN and STUMP cases. Loss of inhibitory function of wild type P53 gene in leiomyosarcoma is an essential event that discriminate frankly malignant tumors from STUMP and atypical leiomyoma.

We examined the expression pattern of cyclooxygenase-2 (COX-2) and c-kit in uterine smooth muscle neoplasms and tried to determine the role of these markers in differential diagnosis. Archival tissue from 64 patients with uterine smooth muscle neoplasms (20 leiomyomas (LMs), 22 atypical leiomyomas (ALMs), and 22 leiomyosarcomas (LMSs)) was immunostained with antibodies against estrogen (ER) and progesterone receptors (PR), COX-2 and c-kit. 7 of 20 LM cases and 5 of 22 ALM cases were immunopositive for COX-2, whereas none of the LMS cases stained immunopositive (p< or =0.05). 4 of 20 LM cases and 5 of 22 ALM cases were immunopositive for c-kit, whereas 15 of 22 LMS cases showed c-kit immunopositivity (p< or =0.05). In conclusion, very few LMs and ALMs show COX-2 immunopositivity. LMSs usually do not express COX-2. COX-2 expression in smooth muscle tumors is not a prominent feature. Therefore, COX-2 inhibitors may not be useful in LMS therapy. C-kit was significantly expressed in uterine LMSs.

Background: Estrogen receptor (ER) positive, lymph node (LN) negative breast cancer usually carries a good prognosis; endocrine therapy is often the only adjuvant treatment. However, a small proportion of such patients do badly (possibly representing those with luminal B cancers identified by gene expression profiling). A clinically applicable method for identifying this subgroup of poor prognosis ER+, LN- patients is required to offer them more intensive adjuvant therapy. Material and methods: A consecutive, unselected series of 1072 new, operable breast cancer patients diagnosed between 2000 and 2004 was studied for ER and progesterone receptor (PR) expression, HER2 status and standard pathological and demographic parameters. ER and PR expression were scored on immunohistochemistry (IHC) on preoperative, diagnostic core biopsies using the “Quickscore” method. This ensured optimal fixation and tissue processing avoiding the variable fixation of resection specimens and the well-described sampling error of tissue micro-array (TMA) methodologies. PR expression is frequently heterogeneous resulting in false negative scores in at least 12% of cases by TMA analysis. HER2 status was assessed using IHC with dual color FISH for cases scoring 2+. Adjuvant therapy was prescribed using standard protocols; all patients with ER+ disease received adjuvant endocrine therapy. Follow up data were obtained from the oncology database, and the registrar of deaths for the date and cause of death. All deaths not attributable to breast cancer were censored at the date of death. Accordingly, the primary end-point was breast cancer-specific overall survival. Survival analysis was carried out by Kaplan-Meier survival curves analysed by the Log-Rank test. Multivariate analysis was carried out using Cox's regression. Results: Overall, PR- cancers had a worse prognosis than PR+ (p&amp;lt;10−12, Hazard Ratio 3.40), even in the ER+ (p=0.006, HR 1.86), LN- (p=10−8, HR=5.33) and LN+ (p=10−11, HR=3.26) sub-groups. In the ER+ LN- group, the absence of PR expression predicted worse prognosis (88% vs 96% 8 year survival, p=0.0003) with a hazard ratio of 3.75. This is considerably more significant than Ki67 IHC scoring reported for a similar group of patients in other studies (HR 2.22). Multivariate analysis demonstrated that PR expression was an independent prognostic variable second only to LN status and more powerful than ER. Discussion: Patients with ER+, LN-, PR- breast cancers have a significantly worse prognosis than those with ER+, LN-, PR+ cancers. Unlike Ki67, PR IHC uses a simple, cost effective, standard methodology (as for ER) and should identify patients who may require chemotherapy in addition to endocrine therapy in this group of otherwise good prognosis patients. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-02-06.