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Carmen Behrens
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OA03 - Advances in Lung Cancer Pathology (ID 897)
- Event: WCLC 2018
- Type: Oral Abstract Session
- Track: Pathology
- Presentations: 1
- Moderators:
- Coordinates: 9/24/2018, 10:30 - 12:00, Room 205 BD
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OA03.05 - Characterization of the Immunologic Intra-Tumor Heterogeneity in Early Stages of Non-Small Cell Lung Cancer by Multiplex Immunofluorescence (ID 13334)
11:15 - 11:25 | Author(s): Carmen Behrens
- Abstract
- Presentation
Background
Recurrence of non-small cell lung carcinoma (NSCLC) is associated with genetic and epigenetic intra-tumor heterogeneity (ITH). The interaction between malignant cells, stromal cells, and tumor-associated immune-cells (TAICs), such as T-cell lymphocytes (TCLs) and tumor-associated macrophages (TAMs), is important for progression of NSCLC and the characterization of the immunologic ITH might be relevant to predict recurrence in surgically treated patients at early stages of NSCLC. The aim of this study was to characterize the immunologic ITH of primary NSCLC tumors at early stages using image analysis and multiplex immunofluorescence (mIF) approaches.
a9ded1e5ce5d75814730bb4caaf49419 Method
Eight cases of stage IA and 8 cases of stage IB surgically resected NSCLC (11 adenocarcinomas, ADCs; and 5 squamous-cell carcinomas, SCCs) with a history of early recurrence were selected for this preliminary analysis. FFPE blocks were obtained and consecutive sections were stained with two panels of mIF for immune profiling, panel 1: pan-cytokeratin (AE1/AE3), PD-L1, PD-1, CD3, CD8, and CD68; panel 2: AE1/AE3, CD3, CD8, granzyme-B (GB), CD45RO, and FOXP3. Three not adjacent, intra-tumor regions (3mm2 each) per case were randomly selected after gridding the whole tumor section. A total of 41 intra-tumor regions were scanned by Vectra multispectral-microscope and analyzed using InForm-software. TAICs were quantified in epithelial and stromal compartments from each intra-tumor region. G-Cross AUC (area under the curve) was computed for specific intervals of distances between TAICs and malignant cells. Median distance between TAICs and malignant cells within each region was calculated.
4c3880bb027f159e801041b1021e88e8 Result
The median density of TCLs and TAMs were 1527 cells/mm2 and 635 cells/mm2, respectively, without significant differences between histologic subtypes. TCLs were predominantly concentered in stromal compartment (median, 2222 cells/mm2) compared with epithelial compartment (median, 332 cells/mm2). Percentage and density of TCLs and TAMs varied 4 and 8 times, respectively, between cases and regions. Non-cytotoxic T-cells and inactive cytotoxic T-cells were the most prevalent phenotypes. Higher density of TAMs and antigen-experienced TCLs were observed in stage IB than stage IA.
8eea62084ca7e541d918e823422bd82e Conclusion
Characterization of immunologic ITH of NSCLC is able by mIF and image analysis with FFPE tumor tissue. There is variability of TAICs densities between regions from the same tumor and different subpopulations were observed. TAMs and exhausted T-cells were more prominent in stage IB (tumor >3cm) suggesting these cells may play an important role in recurrence. Ongoing studies with a larger cohort and comparison with non-recurrent surgically treated patients are warranted. Supported by CPRITRP160668 and UTLungSPORE grants
6f8b794f3246b0c1e1780bb4d4d5dc53Only Members that have purchased this event or have registered via an access code will be able to view this content. To view this presentation, please login, select "Add to Cart" and proceed to checkout. If you would like to become a member of IASLC, please click here.
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P1.03 - Biology (Not CME Accredited Session) (ID 935)
- Event: WCLC 2018
- Type: Poster Viewing in the Exhibit Hall
- Track:
- Presentations: 1
- Moderators:
- Coordinates: 9/24/2018, 16:45 - 18:00, Exhibit Hall
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P1.03-24 - TMPRSS4: A Novel Prognostic Biomarker and Therapeutic Target in NSCLC (ID 11988)
16:45 - 18:00 | Author(s): Carmen Behrens
- Abstract
Background
Genomic analyses are identifying novel genes involved in the pathogenesis of non-small cell lung cancer (NSCLC). TMPRSS4, a membrane-anchored serine protease, was previously found as highly overexpressed in NSCLC. Since proteases have been functionally related to cancer growth and metastasis, we sought to study the prognostic value and role of TMPRSS4 in NSCLC.
a9ded1e5ce5d75814730bb4caaf49419 Method
TMPRSS4 expression was evaluated by immunohistochemistry and H-score calculation in TMAs containing a total number of 455 cases. Kaplan-Meier, log-rank and Cox analyses were used to study the prognostic value. In addition, functional assays using NSCLC cell lines and in vivo models were used to assess the possible role of this protease in NSCLC.
4c3880bb027f159e801041b1021e88e8 Result
High expression of TMPRSS4 was associated with reduced relapse-free survival (RFS, p=0.003) and overall survival (OS, p=0.007) in NSCLC patients. The prognostic value was also found in patients with stages I-II. Multivariant Cox regression analysis identified TMPRSS4 as an independent prognostic factor in NSCLC for both RFS (HR 1.61 [1.16-2.23], p<0.004) and OS (HR 1.52 [1.14-2.03], p<0.005). In functional studies we developed genetic systems to overexpress or reduce TMPRSS4 levels in lung cancer cells lines. Overexpression in LKR13 cells led to increased clonogenicity, migration and multiorganic metastasis in liver, bone and suprarenal gland. Abrogation of TMPRSS4 in H358 and H2170 cell lines caused a very strong reduction in proliferation (>70%, 96h after plating), clonocenicity (>90%, after 15 days in culture) and subcutaneous tumor growth. Reduction in S and G2/M phases of the cell cycle, increased apoptosis, and changes in gene expression of cell replication- and migration-promoting genes (i.e. MCM6, TYMS and CDKN1A(p21)) were also found. Cells lacking TMPRSS4 were highly sensitized to chemotherapy, including cisplatin, paclitaxel and gemcitabine, which significantly enhanced the antiproliferative, antitumor and proapoptotic effect of these drugs.
8eea62084ca7e541d918e823422bd82e Conclusion
Our results show that TMPRSS4 is a biomarker of poor prognosis in NSCLC and plays an important role in tumor growth and metastasis, and suggest that its blockade may enhance sensitivity to chemotherapy.
6f8b794f3246b0c1e1780bb4d4d5dc53
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P1.04 - Immunooncology (Not CME Accredited Session) (ID 936)
- Event: WCLC 2018
- Type: Poster Viewing in the Exhibit Hall
- Track:
- Presentations: 1
- Moderators:
- Coordinates: 9/24/2018, 16:45 - 18:00, Exhibit Hall
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P1.04-22 - CD73 Immunohistochemical Expression in Malignant Cells and Correlation with Immune Infiltrate in Non-Small Cell Lung Carcinoma (NSCLC). (ID 11859)
16:45 - 18:00 | Author(s): Carmen Behrens
- Abstract
Background
CD73 is potential novel target for lung cancer immunotherapy involved in the adenosine pathway that induces tumor microenvironment immunosuppression. We investigated the immunohistochemical (IHC) expression of CD73 in a large cohort of NSCLC and correlated with tumor’s clinical, pathological, molecular and immune cells infiltration data.
a9ded1e5ce5d75814730bb4caaf49419 Method
We examined 175 surgically resected stages I-III formalin-fixed and paraffin-embedded NSCLC tumors tissue microarrays, including 107 adenocarcinomas (ADC) and 68 squamous cell carcinomas (SqCC). For IHC, we used the anti-CD73 antibody (clone D7F9A, Cell Signaling Technology) and evaluated membrane (basolateral and luminal) expression in malignant cells. In a subset of cases, CD73 IHC expression was correlated with data available on: a) CD73 gene mRNA expression (Illumina arrays; n=91); b) EGFR and KRAS mutation status and mutational load (whole exome sequencing; n=104); and, c) density of tumor associated immune cells infiltration (CD3, CD4, CD8, CD68, CD45RO, CD57, FOXP3, and granzyme B) and immune checkpoints expression (PD-L1, PD-1, ICOS, TIM-3, IDO-1, B7-H3, B7-H4, VISTA and OX40) assessed by IHC and image analysis (n=172).
4c3880bb027f159e801041b1021e88e8 Result
ADC showed higher CD73 IHC expression than SqCC (P<0.0001). Pathological stage I ADCs showed higher CD73 expression than higher tumor stages (P=0.0419). Using any level of CD73 expression (>1%) CD73 was expressed in 73% and 40% of ADCs and SqCs, respectively. High expression (>50% of malignant cells) was detected in 35% of ADCs and 20% of SqCC. No other significant correlations with clinical-pathological variables, including patients’ outcome were found. Interestingly, ADCs with EGFR (P=0.04) and KRAS (P=0.02) mutation expressed higher CD73 levels than wild-type tumors. In ADC, CD73 IHC expression correlated significantly with the density of immune T CD3+, CD4+, CD8+, CD45RO+ and FOXP3+ cells, as well as macrophages CD68+ cells in tumors (r values range: 0.22-0.45; P values range: 0.001-0.02). Overall, we did not find significant correlations between CD73 immunostaining and the IHC expression of the immune checkpoints examined. CD73 IHC expression correlated positively with mRNA CD73 gene expression levels in all NSCLCs (r=0.6; P<0.0001), ADCs (r=0.6; P<0.0001), and SqCCs (r=0.49, P<0.0001) histology
8eea62084ca7e541d918e823422bd82e Conclusion
We identified that CD73 protein expresses in a subset of resected NSCLCs, being significantly higher in adenocarcinoma histology. In this histology type, CD73 correlates with immune T cells and macrophages infiltration, and notably, with tumor’s EGFR and KRAS mutation. Our data suggest that CD73 is a potential target for NSCLC, particularly for adenocarcinoma histology (Supported by grants CPRIT RP160668 and UT Lung SPORE).
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