Percentage Of Tumor Area Research Articles

EXPRESSION OF ANDROGEN RECEPTOR IN BREAST CANCER BRAIN METASTASES

Abstract Given availability of central nervous system (CNS)-penetrant systemic therapies that target the androgen receptor (AR), we evaluated the expression of the AR “target” in breast cancer brain metastases (BrM). METHODS: An established, retrospective cohort of 57 patients with metastatic breast cancer who underwent surgery for BrM at the Sunnybrook Odette Cancer Centre (SOCC) between 1999 and 2013 was studied. AR expression in BrM samples was assessed in triplicate using immunohistochemistry (IHC). AR positive status was defined as nuclear AR expression ≥10% in tumor-infiltrating cells as a percentage of tumor area using the SP107 antibody. RESULTS: The median age of patients was 52 years (range 32-85 years). 17 (30%) patients had hormone receptor positive (HR+) /HER2-negative, 28 (49%) had HER2+, and 12 (21%) had triple negative breast cancer (TNBC) BrM. The median expression of AR was 20% (CI 1.6-38.3%) and 32 of 57 (56%) BrM were AR positive based on a cut-point of ≥10%. A significantly smaller proportion of patients with TNBC had AR+ BrM (n=2/12, 17%), compared to patients with HR+/HER2-ve (n= 9/17, 53%) or HER2+ (n=21/28, 75%) disease (p=0.04). Patients with AR positive versus AR negative BrM had similar overall survival (12.5 vs. 7.9 months, p= 0.6), brain-specific progression-free survival (8.0 vs. 5.1 months, p= 0.95), and time from breast cancer diagnosis to BrM diagnosis (51 vs. 29 months, p=0.16). CONCLUSION: AR is expressed in the majority of breast cancer BrM and represents a promising therapeutic target.

Tumor-Infiltrating Lymphocytes and PD-L1 Expression in Pleomorphic Lobular Breast Carcinoma.

The prognostic and predictive role of stromal tumor-infiltrating lymphocytes (sTILs) is undetermined in pleomorphic invasive lobular cancer (pILC). The same applies for the expression of PD-1/PD-L1 in this rare breast cancer subtype. Here, we aimed to investigate the expression of sTILs and analyze the PD-L1 expression levels in pILC. Archival tissues from sixty-six patients with pILC were collected. The sTIL density was scored as a percentage of tumor area using the following cut-offs: 0%; <5%; 5-9%; and 10-50%. The PD-L1 expression was analyzed using IHC on formalin-fixed, paraffin-embedded tissue sections using SP142 and 22C3 antibodies. A total of 82% of the sixty-six patients were hormone receptor positive and 8% of cases were triple negative (TN), while 10% showed human epidermal growth factor receptor 2 (HER2) amplification. sTILs (≥1%) were present in 64% of the study population. Using the SP142 antibody, 36% of tumors demonstrated a positive PD-L1 score of ≥1%, and using the 22C3 antibody, 28% had a positive PD-L1 score of ≥1. There was no correlation between sTILs or PD-L1 expression and tumor size, tumor grade, nodal status, expression of estrogen receptor (ER), or amplification of HER2. Our data did not show any difference in survival between the three molecular subtypes of pILC with respect to sTILs and PD-L1 expression. This study shows that pILCs show some degree of sTILs and PD-L1 expression; however, this was not associated with a survival improvement. Additional large trials are needed to understand immune infiltration in lobular cancer, especially in the pleomorphic subtype.

PD-L1 immune cell expression levels and clinical outcomes in advanced triple-negative breast cancer treated with atezolizumab.

e13066 Background: Treatment with immune checkpoint inhibitors (ICI) has become a standard of care for metastatic PD-L1 positive TNBC patients and PD-L1 has emerged as an important predictive biomarker. Predictive differences of PD-L1 were found in response to immunotherapy in different studies. Alterations in DNA repair pathways (aDDR) have also been investigated as a predictive biomarker for ICI due to the increased mutations and neoantigen production. Real-world data regarding outcomes of treatment of metastatic (m)TNBC patients (pts) with ICI and predictive value of PD-L1 and aDDR is lacking. In this study PD-L1 and aDDR were evaluated for association with clinical benefit of treatment with atezolizumab and nab-paclitaxel (A+nP) in PD-L1 positive mTNBC patients treated in routine clinical practice. Methods: Medical records from 41 PD-L1 positive mTNBC patients consecutively treated with A+nP at Oncology Institute Ljubljana between April 2019 and October 2021 were collected for the present study. PD-L1 expression was assessed on immune cells (IC) as percentage of tumor area using VENTANA SP142 PD-L1 IHC assay (&lt;1% [PD-L1 negative] vs. ≥1% [PD-L1 positive). Patients were divided into 2 subgroups based on PD-L1 status, high (PD-L1≥ 5%) and low (PD-L1 ≥1% and &lt;5%). Germline alteration status in DDR genes was determined on blood samples. We used Kaplan-Meier survival curves to calculate median (m) PFS and OS . The prognostic impact of PD-L1exspresion levels and aDDR was calculated using Cox model. Results: Median follow up of the study was 22 months. Majority of patients had low PD-L1 status (25/41, 61%) and 54% of evaluable patients (15/28) were found to harbor deleterious alterations in DDR genes, of them BRCA1 in 9 pts, BRCA2 in 2 pts, and PALB2, CHEK2, CHD1, RAD51D in 1 pts. Median PFS in the entire cohort was 8.26 months; 95% CI (3.54-12.98) and mOS 20.49 months; 95% CI (11.3-29.7). Significant survival benefit was observed among patients with high PD-L1 status treated with A+nP. No difference in survival was found in patients harbouring germline mutations (aDDR positive) versus without mutations (Table). Conclusions: PD-L1 positive mTNBC patients with high PD-L1 status (≥5%) had almost three times less risk of progression and death compared to low PD-L1 status (≥1% and &lt;5%). Presence of aDDR showed no effect on disease outcome. [Table: see text]

Abstract P5-13-17: PD-L1 expression in breast to brain metastases

Abstract Background Brain metastases (BrM) are a major cause of morbidity and mortality in women with breast cancer. Immunotherapy has the potential for intracranial efficacy among patients with breast cancer since intracranial response to immunotherapy has been observed among patients with other solid tumors. The aim of the study is to analyze the immunohistochemical expression of programmed death ligand 1 (PD-L1), a predictive biomarker of response to immunotherapy, in breast cancer BrMs. Methods A retrospective cohort study of consecutive patients who underwent surgery for breast cancer BrM at Sunnybrook Health Sciences Centre between July 1999 and June 2013 were identified through the Anatomic Pathology departmental database. A tissue microarray using 1um cores was obtained. Immunohistochemical expression of PD-L1 in BrM tissue was assessed in triplicate; PD-L1 positive status was defined as PD-L1 expression ≥1% in tumor infiltrating cells as a percentage of tumor area using the Ventana SP142 antibody. Estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor (HER2) status was determined using 2018 ASCO/CAP guidelines. Results The median patient age at the time of BrM diagnosis was 52 (range 32-85). ER, PR and HER2 status was available in 58 of 59 cases as follows: 12 (20.3%) triple negative (TNBC), 14 (23.7%) HER2+/HR-, 14 (23.7%) HER2+/HR+, 18 (30.5%) hormone receptor (HR)+/HER2-. The majority 62.7% (n=37) of patients had a solitary BrM. The median size of BrM was 2.9 cm (range 0.3 cm to 6.2 cm) with the most common location being the cerebellum and frontal lobe. The majority of patients (n=51, 86.4%) had symptomatic BrM. The most common sites of extra-cranial metastases were bone (37%), lung (32%), liver (22%), lymph nodes (18.6%), and chest wall (3.4%). After surgical excision of BrM, 88.1% of patients received adjuvant stereotactic radiosurgery (SRS) or whole brain radiotherapy (WBRT). 39% of patients received at least 1 line of systemic therapy for metastatic disease prior to the development of BrM. The median follow-up for BrM events was 16.1 months. PD-L1 status was available for all 59 patients. In the overall cohort, 9 out 59 (15.3%) breast cancer BrM were PD-L1 positive irrespective of subtype. The frequency of PD-L1 positive BrM by subtype is as follows: TNBC (n= 3/12, 25%), HER2+/HR- (n=3/14, 21.4%), HER2+/HR+ (n=1/14, 7.1%), and HR+/HER2- (n=2/18, 11.1%). Expression of PD-L1 in BrMs was not associated with patient age at the time of BrM diagnosis, size or location of the BrM, grade of the BrM nor breast cancer subtype. At 24 months, brain-specific progression-free survival (bsPFS) was 47.5% (95% CI 32.9-68.7%). When stratified by PD-L1 status, 24-month bs-PFS was 66.7% (95% CI 37.9-100%) among patients with PD-L1 positive BrM versus 42% (95% CI 26.6-67.3%) among those with PD-L1 negative BrM (log-rank p-value 0.142). Conclusion One in 7 patients in our cohort had PD-L1 positive BrM; this proportion was highest (25%) among those with TNBC. Hence, there is rationale to include patients with breast cancer BrM in clinical trials evaluating efficacy of immunotherapy. Citation Format: Rania Chehade, Maleeha A Qazi, Marguerite Ennis, Sharon Nofech-Mozes, Katarzyna Jerzak. PD-L1 expression in breast to brain metastases [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-13-17.

PD-L1 expression in breast cancer brain metastases.

BackgroundTo evaluate the potential intracranial efficacy of immunotherapy among patients with breast cancer brain metastases (BrM), we analyzed the immunohistochemical expression of programmed death-ligand 1 (PD-L1), a predictive biomarker of response to immunotherapy.MethodsIn this single-center retrospective cohort study, consecutive patients with breast cancer BrM (immunotherapy naïve) who underwent surgery for BrM at Sunnybrook Health Sciences Center between July 1999 and June 2013 were identified. PD-L1 expression by immunohistochemistry (IHC) was assessed on BrM samples in triplicate; PD-L1 positive status was defined as PD-L1 expression ≥1% on tumor-infiltrating cells as a percentage of tumor area using the Ventana SP142 antibody. Estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor (HER2) status was determined using 2018 ASCO/CAP guidelines.ResultsThe median patient age at the time of BrM diagnosis was 52 (range 32–85). PD-L1 expression using the SP42 antibody was identified in 9 out of 59 (15.3%) breast cancer BrM. The frequency of PD-L1 positive BrM by subtype is as follows: TNBC (n = 3/12, 25.0%), HER2+/HR- (n = 3/14, 21.4%), HR+/HER2- (n = 2/18, 11.1%), and HER2+/HR+ (n = 1/14, 7.1%). 24-month brain-specific progression-free survival was 66.7% (95% CI 37.9%–100%) among patients with PD-L1 positive BrM versus 42% (95% CI 26.6%–67.3%) among those with PD-L1 negative BrM (log-rank P-value .142).ConclusionsOne in 7 patients in our cohort had PD-L1 positive BrM; this proportion was highest (25%) among those with TNBC. Intracranial efficacy of immunotherapy warrants further study, particularly among patients with treatment-naïve metastatic TNBC, for whom extracranial efficacy has already been established.

Transient Competitive Inhibition Bypasses the Binding Site Barrier to Improve Tumor Penetration of Trastuzumab and Enhance T-DM1 Efficacy.

Poor penetration of mAbs in solid tumors is explained, in part, by the binding site barrier hypothesis. Following extravasation, mAbs rapidly bind cellular antigens, leading to the observation that, at subsaturating doses, therapeutic antibody in solid tumors localizes around tumor vasculature. Here we report a unique strategy to overcome the binding site barrier through transient competitive inhibition of antibody-antigen binding. The anti-trastuzumab single domain antibody 1HE was identified through in vitro binding assays as a model inhibitor. Coadministration of 1HE did not alter the plasma pharmacokinetics of trastuzumab or ado-trastuzumab emtansine (T-DM1) in vivo. Administration of 1HE alone was rapidly eliminated with a terminal plasma half-life of 1.2 hours, while coadministrations of 1HE with trastuzumab had a terminal half-life of 56 hours. In mice harboring SKOV3 xenografts, coadministration of 1HE with trastuzumab led to significant increases in both penetration of trastuzumab from vasculature and the percentage of tumor area that stained positive for trastuzumab. 1HE coadministered with a single dose of T-DM1 to NCI-N87 xenograft-bearing mice significantly enhanced T-DM1 efficacy, increasing median survival. These results support the hypothesis that transient competitive inhibition can improve therapeutic antibody distribution in solid tumors and enhance antibody efficacy. SIGNIFICANCE: This study describes the development of a transient competitive inhibition strategy that enhances the tumor penetration and efficacy of anticancer antibodies.See related commentary by van Dongen, p. 3956.

Early Surgical Treatment of Both Tumor and Fracture in Patients With Enchondroma of the Hand Combined With Pathologic Fracture.

The aims of this study were to evaluate the degree of cortical thinning and the tumor area in simple radiographs in patients with enchondroma of the hand combined with pathologic fracture and to determine the surgical outcome of simultaneous tumor curettage and internal fixation of fracture. We enrolled 23 bone lesions of 21 patients diagnosed with enchondroma of the hand. Of them, 9 bone lesions of 8 patients had pathologic fracture. We evaluated the degree of cortical thinning and tumor area in simple radiographs and compared them between patients with and without pathologic fracture. For patients with pathologic fracture, we performed curettage of the tumor through the fracture site and fixed the fracture using K-wires. We then evaluated the clinical outcome of these patients using the visual analog scale to measure pain intensity and Takigawa's criteria 1 year after surgery. There was a significant difference in the degree of cortical thinning between the 2 groups (P < 0.001); however, there was no significant difference in the percentage of tumor area (P = 0.259). The average time from surgery to bony union was 8.5 weeks in patients with pathologic fracture. The patients reported that they could return to their previous activities of daily living within 12 weeks of surgery. The average visual analog scale score was 0.6 at 1 year after surgery, and 6 patients were graded as excellent and 2 as good according to Takigawa's criteria. Cortical thinning seems to be related to pathologic fracture. Our scoring system for cortical thinning may be helpful in evaluating the risk for pathologic fracture. Early 1-stage surgical treatment for pathologic hand fracture with enchondroma is a very useful method with satisfactory outcomes that could shorten the treatment period.

P2.09-15 PD-L1 Expression and Lymphocyte Infiltration in Resected Stage IIIAN2 NSCLC: Preliminary Data from a Lung ART Ancillary Study

Patients with resectable stage IIIA N2 NSCLC, are at high risk of both systemic and loco-regional relapse following surgical resection, necessitating neo-adjuvant or adjuvant treatments. Prognostic biological markers are needed. Parameters from the immune microenvironment, including PD-L1 expression and lymphocytic infiltration, have been poorly described in this group of patients. Thus we assessed simultaneously PD-L1 expression and TIL density in a cohort of stage IIIA N2 Lung ART patients, and correlated the results with clinical and pathological features before adjuvant treatment. Formalin fixed paraffin-embedded tumor surgical specimens from 247 patients included in the Lung Adjuvant Radiotherapy Trial (NCT00410683) were studied. PD-L1 immunohistochemistry was performed centrally on whole slides using a validated clinical PD-L1 assay. Expression of PD-L1 in tumor cells (TC) and immune cell (IC) was scored by a trained pathologist. Morphological assessment of TIL density (percentage of tumor area) was performed on whole hematoxylin-eosin stained slides. Surgical and pathology reports were reviewed by an independent expert committee for tumor staging. Association between immune parameters and baseline clinical characteristics were assessed in exploratory analyses in order to provide insights on immune activity in resected NSCLC patients. PD-L1 expression in ≥1% TC, ≥50% TC, ≥1% IC, ≥10% IC was observed in 47.8%, 21.9%, 61.5%, 7.3% of patients, respectively. In univariate analysis, high PD-L1 expression in both tumor cells and immune cells for all cut points correlated strongly with a higher TIL density (p-values ≤0.001). In 41 (16.6%) patients with preoperative chemotherapy (CT), a higher TIL density was observed (mean 28.1 vs. 17.5%, p=0.0018) as compared to patients without preoperative CT, but no difference was noted for PD-L1 expression in both TC and IC,. Skip N2 metastases were associated with a higher TIL infiltration (mean 22.9% vs. 17.4% p=0.014). We found no significant correlation between PD-L1 or TIL infiltration with the number of mediastinal lymph nodes stations involved on pathological examination and with histological tumor subtypes (squamous cell carcinoma vs. adenocarcinoma). PD-L1 expression levels in TC and IC appeared similar in stage IIIA N2 NSCLC as compared to other stages. Expression in both TC and IC strongly correlated with TIL infiltration, suggesting a prominently immune-induced expression mechanism. Preoperative chemotherapy was associated with a higher TIL infiltration but not higher PD-L1 expression. Patients with skip N2 metastases harbored a higher level of TIL density, a finding consistent with a more active immune microenvironment in this group of patients with better prognosis. These data will be subsequently updated on a larger number of patient and correlated to clinical follow-up.

Abstract PD6-01: Prevalence of PDL1 and tumor infiltrating lymphocytes (TILs) in primary and metastatic TNBC

Abstract Background: Metastatic Triple Negative Breast Cancer (TNBC) is an aggressive breast cancer subtype with a high unmet medical need and limited therapeutic options. PDL1 on immune cells (IC) and TILs were associated with activity to the anti-PDL1 mAB atezolizumab in mTNBC (Schmid AACR 2017). PDL1 IC &amp;gt;=1% is the diagnostic hypothesis for TNBC atezolizumab studies. The goal of the current project was to evaluate the prevalence of PDL1 and TILs in primary and metastatic TNBC samples. Methods: FFPE tumor tissue from pathology documented TNBC tissues were centrally tested for PDL1 IHC on tumor cells (TC) and infiltrating immune cells (IC) (VENTANA SP142 assay) and quantified as percentage of tumor area. TILs were evaluated as a percentage of tumor area and PD-L1 gene (CD274) expression was quantified by Fluidigm. Results: 669 tumors were evaluable for PD-L1 by IHC and TILs. 280 samples were annotated as primary tumors and 179 as metastases with 210 denoted as unknown. PDL1 IC and TC immunostaining was correlated to PD-L1 transcript expression (r = 0.55 and r = 0.51, respectively). 84% of samples had no tumor cells expressing PDL1 and 16% had at least 1% of PDL1 in TC. 44% of the samples had no PDL1 in the immune cells, 35% expressed PDL1 IC [31% to &amp;lt;5%] and 21% PDL1 IC 35%. PDL1 distribution was similar between primary and metastases, with median of 1% and 0% for PDL1 IC and TC, respectively. Analyses of anatomical locations suggest that PDL1 IC is highest in lymph nodes (median IC = 3%), lowest in liver metastases (median IC = 0.5%) while breast tissue is intermediate (median IC = 1%). The correlation between PDL1 IC and TILs is r = 0.69. TILs were numerically higher in primary compared to metastases (10% vs 5%). Anatomical TIL distribution confirmed liver metastases as least infiltrated, while breast, lymph node and lung metastases with similarly higher values (5% vs 10%, respectively). Concordance analyses in intra-individual synchronous and asynchronous matched pairs were performed at a cutoff of PDL1 IC 31%. Concordance of matched specimens was 82% in 11 synchronous pairs and 75% in 20 asynchronous pairs. Conclusions: Prevalence of PDL1 IHC is similar between primary and metastatic TNBC samples. Although PDL1 IC and TILs are highly correlated, TILs are less prevalent in metastases compared to primary tissue. Anatomical location may influence prevalence of both PDL1 IC and TILs. Concordance of PDL1 in matched pairs is high, suggesting PDL1 IC may be a stable biomarker. Citation Format: Li Y, Chang C-W, Tran D, Denker M, Hegde P, Molinero L. Prevalence of PDL1 and tumor infiltrating lymphocytes (TILs) in primary and metastatic TNBC [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr PD6-01.

PD-L1 expression and the immune microenvironment in primary invasive lobular carcinomas of the breast

Tumor-infiltrating lymphocytes and immune checkpoint proteins such as PD-L1 are potential prognostic factors and therapeutic targets in breast cancer. Most studies characterizing the breast tumor immune microenvironment have focused on ductal carcinomas. Here we investigate the tumor microenvironment of primary invasive lobular carcinomas. Previously constructed tissue microarrays of 47 lobular carcinomas were labeled by immunohistochemistry for PD-L1, CD8, CD20, and FoxP3. The stromal immune infiltrate density was qualitatively scored as a percentage of tumor area: 1+ (<5%); 2+ (5–10%); 3+ (10–15%); or 4+ (>50%). The average immune cell subtype per high-power field was quantitatively scored. The percentage PD-L1 labeling on tumor-infiltrating lymphocytes was scored as none, focal (<5%), moderate (10–24%), or diffuse (50–100%). The percentage of membranous carcinoma cell PD-L1 labeling was also recorded, with <5% considered negative. All lobular carcinomas contained PD-L1+ tumor-infiltrating lymphocytes with the majority showing 1+ immune infiltrates with focal–moderate PD-L1 labeling. PD-L1 was expressed by tumor cells in 17% of lobular carcinomas. In contrast to ductal carcinomas, there was no correlation between the immune infiltrate density, the PD-L1 expression by lobular carcinoma cells, tumor grade, or the expression of estrogen receptor or human epidermal growth factor receptor-2. However, both the tumor-infiltrating lymphocyte density and the average CD8+ T-cell counts correlated with immune cell PD-L1 status (P=0.004 and 0.03, respectively). Similar to breast ductal carcinomas, PD-L1+ lobular breast carcinomas had higher numbers of PD-L1+ tumor-infiltrating lymphocytes (63%) than PD-L1− lobular carcinomas (23%; P=0.04). These data show that a subset of primary breast lobular carcinomas both express PD-L1 on tumor cells and contain PD-L1+ tumor-infiltrating lymphocytes, suggesting the possibility of both constitutive and adaptive PD-L1 expression. Together, these results support immunotherapy as a potential treatment for a subset of patients with primary invasive lobular breast carcinomas.

Investigation of hypoxia conditions using oxygen-enhanced magnetic resonance imaging measurements in glioma models.

The objective of this study was to determine whether using oxygen-enhanced magnetic resonance imaging (OE-MRI) to assess hypoxia is feasible and whether historical measurements, pO2 changes, and percentage of signal intensity changes (PSIC) are correlated in an animal model of glioma. A total of 25 Sprague-Dawley rats were used to establish C6 brain or subcutaneous glioma model. Nine rats with brain gliomas underwent OE-MRI followed by histopathologic analysis to assess microvessel density and hypoxia. Another 11 rats were underwent OE-MRI and were followed for a survival analysis. Time–T1-weighted MR signal intensity (SI) curves and PSIC maps were derived from the OE-MRI data. High–regions of interests (ROI-h; PSIC > 10%) and low-ROIs (ROI-l; PSIC < 10%) were defined on the PSIC maps. To validate the PSIC map for identifying tumor hypoxia, we subjected an additional 5 rats with subcutaneous glioma to OE-MRI and pO2 measurements. All tumors showed regional heterogeneity on the PSIC maps. For the brain tumors, the time-SI curves for the ROIs-h showed a greater increase in SI than those for the ROIs-l did. The percentage of tumor area with a low PSIC was significantly correlated with the percentage of hypoxia staining and necrosis (r =0.71; P<0.05). ROIs with a higher PSIC typically had more vessels (r=0.88; P<0.05). A significant difference in survival was shown (log-rank P = 0.035). The time-pO2 curves of the subcutaneous tumors were similar to the time-SI curves. PSIC was significantly correlated with pO2 changes (r =0.82; P<0.05). These findings suggest that OE-MRI measurements can be used to assess hypoxia in C6 glioma models. In these models, the PSIC value was correlated with survival, indicating that PSIC could serve as a prognostic marker for glioma.

1171P - The SP142 PD-L1 IHC assay for atezolizumab (atezo) reflects pre-existing immune status in NSCLC and correlates with PD-L1 mRNA

PD-L1 expression as measured by the SP142 PD-L1 IHC assay is associated with improved efficacy with atezo in patients with NSCLC, including overall survival. In capturing PD-L1 on both tumor cells (TC) and tumor-infiltrating immune cells (IC), this assay is able to determine preexisting immune status, underscoring the predictive value of PD-L1 IHC. In this study, we further investigated intratumoral immune infiltration status by measuring T effector (Teff) gene signature and correlated PD-L1 mRNA expression with SP142 PD-L1 IHC. Using the SP142 IHC assay, procured samples from NSCLC tumors were scored according to the percent of TC expressing PD-L1 (TC0 < 1%; TC1 ≥ 1% and < 5%; TC2 ≥ 5% and < 50%; TC3 ≥ 50%). IC were scored as percentage of tumor area (IC0 < 1%; IC1 ≥ 1% and < 5%; IC2 ≥ 5% and < 10%; IC3 ≥ 10%). Four mutually exclusive groups increasing in PD-L1 expression (TC0 and IC0, TC1 or IC1, TC2 or IC2, TC3 or IC3) were designated. mRNA levels for PD-L1 and Teff signature were measured in the same NSCLC specimens using the Fluidigm platform and correlated with each of the PD-L1 IHC sub-groups. In 86 NSCLC specimens, mRNA levels for PD-L1 increased incrementally with higher PD-L1 IHC status, from 0 median expression for TC0 and IC0 tumors to 3.14 median expression for TC3 or IC3 tumors. A similar trend was seen for the Teff signature, with values increasing from 0 to 1.65 median expression with increasing IHC cutoffs (Table). Results from the SP142 PD-L1 IHC assay associates with PD-L1 and Teff mRNA transcripts, reflecting pre-existing immunity associated with adaptive PD-L1 expression on IC as well as intrinsic PD-L1 expression on TC in NSCLC, thus providing an independent orthogonal correlation of IHC to gene expression. The value of mRNA gene expression as a predictive marker of efficacy for atezo in NSCLC is being studied in ongoing randomized trials of atezo.Tabled 1PD-L1 IHC vs PD-L1 mRNA and T effector gene signaturePD-L1 IHCPD-L1 mRNAT EffectoranMedianb expression95% CInMedianb expression95% CITC0 and IC0460-0.22, 0.36460-0.39, 0.16TC1 or IC1220.890.53, 1.66220.730.25, 0.95TC2 or IC2112.662.49, 2.85111.530.55, 1.57TC3 or IC373.141.4, 4.2971.650.46, 1.95a Includes CD8A, GZMA, GZMB, EOMES, CXCL9, CXCL10, TBX21. b Median expressions are log scale expression (-ΔCT) relative to the median of TC0 and IC0 group. Larger values indicate higher expression. Open table in a new tab

Tie2-Expressing Monocytes Are Associated with Identification and Prognoses of Hepatitis B Virus Related Hepatocellular Carcinoma after Resection.

BackgroundTie2-expressing monocytes (TEMs) are found in various tumors, involved in forming tumor blood vessels and expressing several important proangiogenic factors. The goals of this study were to evaluate the value of TEMs in diagnosing and predicting the prognosis of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC).MethodsFlow cytometry was performed to identify and count TEMs in peripheral blood monocytes from HCC patients (n = 84) receiving hepatectomy, HBV cirrhotic patients (n = 21), benign tumors patients (n = 15) and healthy volunteers (n = 23). Angiopoietin-2 (Ang-2) levels in the plasma were determined by enzyme linked immunosorbent assay. The distribution of TEMs in tumor tissue was observed by immunofluorescence staining. Then we determined the vascular area as a percentage of tumor area (vascular area/tumor area) by immunohistochemical staining. Finally the prognostic significance of TEMs and other clinicopathologic factors was evaluated.ResultsPercentage of TEMs in peripheral blood monocytes significantly increased in HCC patients compared with HBV cirrhotic patients and healthy donors (both P< 0.001). However there was no significance in benign liver tumor (P = 0.482). In addition, the percentage of circulating TEMs was positively correlated with plasma Ang-2 concentration (P<0.001, r2 = 0.294) and vascular area/tumor area (P<0.001, r2 = 0.126). Furthermore the percentage of intratumoral TEMs was significantly higher than that of paratumoral TEMs (P<0.001). Increased circulating TEMs was associated with poor overall survival (P = 0.043) and a shorter time to recurrence (P = 0.041). Multivariate Cox analysis also revealed that the percentage of TEMs in peripheral blood was an independent factor for HCC patients’ prognosis.ConclusionsTEMs may promote angiogenesis in HCC regarding the angiopoietin/Tie2 signal pathway. Percentage of TEMs in peripheral blood monocytes may be applied as a biomarker for identifying HBV-related HCC and predicting the prognosis of these patients after resection.

Prognostic associations of insulin-like growth factor-1 receptor in primary uveal melanoma

Objective To study the association between immunoreactivity for insulin-like growth factor-1 receptor (IGF-1R) in primary ciliary body and choroidal melanoma and metastatic death in a consecutive, population-based data set. Design Retrospective, consecutive, population-based cohort study. Participants A total of 167 patients with choroidal and ciliary body melanoma, enucleated from 1972 to 1981, with long-term survival data. Methods Specimens were immunostained by using the avidin-biotinylated peroxidase complex method and polyclonal antibodies to IGF-1R. The percentage of tumour area that was immunopositive was recorded. Survival was assessed by Cox multivariate regression analysis. Results The tumour area could be reliably measured from 129 (78%) of the 167 choroidal or ciliary body melanomas. More heavy pigmentation ( p = 0.001), larger number of macrophages ( p = 0.003) and higher microvascular density ( p = 0.060) were associated with a higher percentage of tumour area that was immunopositive for IGF-1R, the reverse being true of extrascleral extension ( p = 0.049). No significant association was observed with ciliary body extension, largest basal tumour diameter, cell type, mean diameter of the 10 largest nucleoli, and presence of extravascular matrix loops and networks ( p = 0.61-0.96). The percentage of tumour area that was immunopositive for IGF-1R was not associated with survival. Conclusions In our data set, immunoreactivity for IGF-IR did not independently predict metastasis from primary uveal melanoma. Partial loss of antigenicity can not be ruled out as a confounding factor because no frozen sections were available. Results of previous studies have likewise been variable, suggesting that immunohistochemical determination of IGF-1R from formalin-fixed, paraffin-embedded specimens is not practical as a routine test.

Inflammatory angiomyolipomas of the liver: a clinicopathologic and immunohistochemical analysis of 5 cases

Hepatic angiomyolipoma (AML) may demonstrate a marked histologic diversity. We report 5 cases of hepatic AML exhibiting prominent inflammatory cells in the background (inflammatory AML). The patients were 4 females and 1 male, with age ranged from 21 to 48 years (mean, 39.2 years). Three tumors were in the left lobe and 2 in the right lobe. The tumor size was from 5.5 to 10 cm in the greatest dimension (mean, 7.46 cm). No patient had clinical features of tuberous sclerosis. Histologically, the striking feature was the infiltration of numerous inflammatory cells in the background of the tumors, including small lymphocytes, plasma cells, and histiocytes. The percentage of tumor area with heavy inflammatory infiltration was more than 50% in all cases. The myoid cells were spindled and epithelioid in shape, with eosinophilic or clear cytoplasm, and were arranged in fascicles and clusters. Scattered adipose cells and sinusoidal and thick-walled blood vessels were variably present in all tumors. Focal trabecular arrangement was present in 2 of the 5 tumors. There was no nuclear atypia, and mitotic figures were rare. The myoid cells were diffusely positive for vimentin, smooth muscle actin, and HMB-45 in all cases. All patients showed no evidence of disease after the initial surgical excision during a follow-up period from 3 to 9 years. The inflammatory AMLs should be distinguished from other tumors with inflammatory background such as inflammatory myofibroblastic tumor and follicular dendritic cell tumor.