bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4563: Profiling the tumor immune microenvironment of adenocarcinoma and squamous cell NSCLC
Authors: Harmeyer Kayla , Deng Jiehui , Shah Suhagi , Bagdatlioglu Ece , Ahsan Aarif , Reid Christopher , Thennavan Aatish , Perou Charles , Wong Kwok-Kin , Pass Harvey
Published: August 2019

The emergence of immune checkpoint inhibitors has changed the treatment paradigm for NSCLC. Recently, immunotherapy in combination with chemotherapy was shown to be a promising first-line treatment strategy for patients with squamous cell lung cancer, where targeted therapy has had little efficacy due to the pathological driver mutations for this disease being poorly understood. However, for both adenocarcinoma and squamous cell NSCLC, use of such immune checkpoint inhibitors has shown to improve the outcome for only a subset of patients with advanced or metastatic disease. Understanding the diversity in the tumor immune microenvironment will be critical to not only advance immune oncology research but to also determine potential clinical distinctions that dictate therapeutic response. Here, we take a multifaceted approach to compare the tumor immune microenvironments of over 50 patients diagnosed with either adenocarcinoma or squamous cell NSCLC. To this end, we performed a comprehensive multicolor flow cytometry analysis to compare tumor immune infiltrates and expression of suppressive markers to adjacent normal lung tissue as well as peripheral blood immune cells from the same patient. This analysis showed differences in the CD4+ and CD8+ T cell, CD19+ B cell, NK cell, and CD14+/CD16+ myeloid cell populations between the tumor and matched tissue and blood. These immune profiles were then compared between adenocarcinoma and squamous cell carcinoma subtypes as well as early versus late stage disease. We then integrated these findings with multiplexed immunofluorescence analysis on a panel of immune cell markers. Finally, 10x genomics single-cell RNA sequencing (scRNAseq) data were used to analyze single-cell transcriptomes from both adenocarcinoma and squamous cell lung tumors. Our result showed great heterogeneity of immune infiltrates across different patient samples, including different subtypes and different disease stages. This detailed comparative analysis will help us further understand the involvement of tumor infiltrating immune cells for different subtypes of lung cancer.

Citation Format: Kayla Harmeyer, Jiehui Deng, Suhagi Shah, Ece Bagdatlioglu, Aarif Ahsan, Christopher Reid, Aatish Thennavan, Charles Perou, Kwok-Kin Wong, Harvey Pass. Profiling the tumor immune microenvironment of adenocarcinoma and squamous cell NSCLC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4563.
Abstract 4577: Anti-tumor immunity is a key determinant of small cell lung cancer survivorship
Authors: Kosari Farhad , Muppa Prasuna , Terra Simone , Sharma Anurag , Mansfield Aaron , Aubry Marie Christine , Janaki Nafiseh , Nasir Aqsa , Peikert Tobias
Published: August 2019

Introduction: While most small cell lung cancer (SCLC) patients die within a few months, a sub-group of patients survive for many years. Factors determining long-term survivorship remain largely unknown. We present the first comprehensive comparative genomic and tumor microenvironment analyses of small cell lung cancer (SCLC) between patients with long term (LTS) and expected (EXS) survival times.

Methods: We compared surgically resected tumors of 23 LTS (survival > 4 years) and 18 EXS (survival ≤ 2 years). There were no differences in clinical variables including TNM staging and curative versus non-curative intend surgery between the groups. Gene expression profiling was performed by microarrays and tumor microenvironment analyses were by IHC of prominent immune related markers.

Results: Immune related genes and pathways represented the majority of the differentially overexpressed genes in LTS compared to the EXS. The differences in the immunological tumor-microenvironment were confirmed by quantitative immuno-staining. Increased numbers of tumor infiltrating and associated lymphocytes were present throughout tumors of LTS. Several differentiating patterns of enhanced anti-tumor immunity were identified. While some areas of LTS tumors also harbored higher numbers of suppressive immune cells (monocytes, regulatory lymphocytes, and macrophages), ratios of these suppressive cells to CD3+ lymphocytes were generally lower in LTS tumors indicating a more tumor suppressive microenvironment.

Conclusions: Our data demonstrate that long-term survivorship of SCLC patients is strongly influenced by the presence of anti-tumor immune cells in the tumor microenvironment. Characterization of the anti-tumor immune responses may identify opportunities for individualized immunotherapies for SCLC.

Citation Format: Farhad Kosari, Prasuna Muppa, Simone Terra, Anurag Sharma, Aaron Mansfield, Marie Christine Aubry, Nafiseh Janaki, Aqsa Nasir, Tobias Peikert. Anti-tumor immunity is a key determinant of small cell lung cancer survivorship [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4577.
Abstract 4580: Primary tumor-induced immunity eradicates disseminated tumor cells in syngeneic mouse model
Authors: Piranlioglu Raziye , Lee Eunmi , Ouzuonova Maria , Rodier Riley , Greer Adam , Bayraktar Feyzanur , Durmus Omer Can , Arbab Ali S. , Thangaraju Muthushamy , Wicha Max S. , Celis Esteban , Korkaya Hasan
Published: August 2019

Although clinically apparent metastasis is associated with late stages of cancer development, micro-metastatic dissemination may be an early event. However, the fate of these early disseminated tumor cells (DTC) remains elusive.

Using the syngeneic mouse models, we demonstrated that although both orthotopically-implanted murine 4T1 and EMT6 tumors are capable of disseminating into secondary organs, only 4T1 tumors develop overt metastasis. However, EMT6 tumors induce an anti-tumor immunity in syngeneic mice and that eradicates disseminated tumor cells (DTC) in distant organs. Following the complete removal of primary EMT6 tumors, mice do not develop detectable metastasis and generate an immunological memory that leads to complete elimination of repeatedly injected tumor cells via tail vein. Conversely, these cells readily grow and metastasize in immuno-deficient athymic or Rag2- mice, and when g-MDSCs from 4T1 tumor-bearing mice were co-injected into immunocompetent EMT6 primed mice. In contrast to complete resection, mice with residual tumors following surgery exhibited an enhanced growth of local and concomitant growth of DTCs at metastatic site with increased g-MDSCs accumulation in lung and spleen.

Together, our results suggest that some tumors are capable of inducing anti-tumor immunity against the DTCs when complete resection of primary tumor cures animals. However, in the presence of residual tumors, inflammation induced by surgical procedure promote the growth of DTCs.

Citation Format: Raziye Piranlioglu, Eunmi Lee, Maria Ouzuonova, Riley Rodier, Adam Greer, Feyzanur Bayraktar, Omer Can Durmus, Ali S. Arbab, Muthushamy Thangaraju, Max S. Wicha, Esteban Celis, Hasan Korkaya. Primary tumor-induced immunity eradicates disseminated tumor cells in syngeneic mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4580.
Abstract 4581: High endothelial venules are associated with immune evasion and hereditary background in mismatch repair-deficient colorectal cancers
Authors: Ahadova Aysel , Pfuderer Pauline , Ballhausen Alexej , Frayling Ian , Ager Ann , Doeberitz Magnus von Knebel , Kloor Matthias
Published: August 2019

DNA mismatch repair (MMR)-deficient cancers can develop in the context of Lynch syndrome or sporadically. MMR-deficient cancers accumulate a high load of immunogenic frameshift peptide neoantigens as a consequence of insertion/deletion mutations at coding microsatellites (microsatellite instability, MSI). Whether enhanced lymphocyte recruitment contributes to the high immune infiltration and the pronounced local immune response typically observed in MSI colorectal cancers (CRCs) is not known. Here we analyzed the density of high endothelial venules (HEVs), postcapillary blood vessels specialized for lymphocyte trafficking, in MSI CRC of sporadic and hereditary origin. Immunohistochemical staining with rat monoclonal MECA-79 antibody was used to detect HEVs in tumor specimens from MSI (hereditary, n=20 and sporadic, n=28) and MSS CRCs (n=35). Lymph follicle-associated HEVs were normalized per mm2 of tumor and peritumoral area. Mutations of the B2M gene were determined by Sanger sequencing. Significantly elevated HEV densities were detected in MSI (n=48) compared to MSS CRCs (n=35) (0.09 vs 0.02 counts per mm2, p=0.0002). Lynch syndrome-associated MSI CRCs presented with significantly higher HEV densities compared to sporadic counterparts (0.15 vs 0.05 counts per mm2, p=0.025). This difference remained significant when age-matched groups of hereditary and sporadic MSI CRCs were compared. MSI CRCs with B2M mutations (n=12) presented with significantly higher HEV densities than their B2M-wild type counterparts (n=14, p=0.016). Our results demonstrate that a high density of HEVs correlates with the MSI phenotype in CRC, underlining the elevated local immune activation in these tumors. Within MSI CRC, Lynch syndrome was a predictor of high HEV counts, suggesting a likely role of lymphocyte recruitment in immune surveillance of Lynch syndrome CRCs. The observation of high HEV counts particularly in MSI CRCs harboring B2M mutations supports the model that immune evasion occurs particularly in an activated local immune environment. The prognostic value of HEVs in MSI CRCs should be addressed in the future studies.

Citation Format: Aysel Ahadova, Pauline Pfuderer, Alexej Ballhausen, Ian Frayling, Ann Ager, Magnus von Knebel Doeberitz, Matthias Kloor. High endothelial venules are associated with immune evasion and hereditary background in mismatch repair-deficient colorectal cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4581.
Abstract 4582: Mannose receptor positive macrophage infiltrate correlates with prostate cancer onset and metastatic castration-resistant disease
Authors: Zarif Jelani C. , Valle Javier A. Baena-Del , Hicks Jessica L. , Heaphy Christopher M. , Vidal Igor , Luo Jacob , Lotan Tamara L. , Hooper Jody E. , Isaacs William B. , Pienta Kenneth J. , Marzo Angelo M. De
Published: August 2019

M2-Tumor Associated macrophages (M2-TAMs) can suppress inflammation within the tumor microenvironment and have been reported to modulate cancer progression. We and others have previously reported infiltration of M2 macrophages in metastatic castrate-resistant prostate cancers (mCRPC). The objective of this study was to determine whether the extent of M2 macrophage infiltration correlates with prostate cancer aggressiveness, we applied immunohistochemistry to normal prostatic tissue, localized prostate cancer and metastatic castrate-resistant prostate cancer (mCRPC) from 192 patients. To assess M2 macrophage involvement, we analytically validated an IHC assay to detect the human mannose receptor (CD206). Also, we used multiplex immunofluorescent staining to show that a small fraction of CD206 staining co-localizes with endothelial cells of lymphatic vessels, while the vast majority of staining occurs in CD68-positive macrophages. The area fraction of staining for CD206-positive macrophages increased in a stepwise fashion going from normal (i.e. non-inflammation) prostatic tissue, to primary untreated carcinomas, to hormone naive regional lymph node metastases to castration resistant prostate cancer. Complimentary studies using flow cytometry confirmed CD206-positive M2-TAM infiltration. Altogether, this study revealed a progressive increase in CD206-positive macrophages from normal prostate to metastatic CRPC. Given the immunosuppressive nature of these cells and lack of clinical success of immunotherapy for prostate cancer patients, our results provide a rationale for therapeutic development to deplete these cells in the prostate cancer microenvironment as a potential method to augment immunotherapeutic approaches in prostate cancer.

Citation Format: Jelani C. Zarif, Javier A. Baena-Del Valle, Jessica L. Hicks, Christopher M. Heaphy, Igor Vidal, Jacob Luo, Tamara L. Lotan, Jody E. Hooper, William B. Isaacs, Kenneth J. Pienta, Angelo M. De Marzo. Mannose receptor positive macrophage infiltrate correlates with prostate cancer onset and metastatic castration-resistant disease [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4582.
Abstract 4593: Inhibition of invasion of HER2-positive breast cancer cells by lysosome targeting drugs
Authors: Hansen Malene B. , Postol Maria , Egan Davig A. , Jäättelä Marja , Kallunki Tuula
Published: August 2019

Up to 20% of all breast cancers are over-expressing HER2/ErbB2, leading to malignant and invasive cancers with a poor patient outcome. Though good ErbB2-targeting treatments have been developed, metastasis formation and drug resistance are still major challenges for the long-term cure. This project aims to find out if some of the numerous pre-existing compounds and clinically approved drugs can be repurposed to treat or improve the treatment of HER2/ErbB2-positive breast cancer. We have therefore set up a high-throughput and high-content screen of the Prestwick Chemical Library to identify drugs that can reverse the ErbB2-induced, invasion-promoting, peripheral distribution of lysosomes. Using a breast cancer cell model expressing N-terminally truncated version of ErbB2, we mimic breast cancer that is resistant to the currently used antibodies, trastuzumab and pertuzumab. This cell line shows a highly invasive phenotype with increased activation of the lysosomal proteases, cysteine cathepsin B and L, as well as anterograde movement of lysosomes. This repositioning of the lysosomes enables them to fuse with the plasma membrane and empty their acidic and hydrolyzing content to the extracellular matrix, promoting invasion. Treatment with the clinically used drug lapatinib will reverse this phenotype. The cells ability to reverse the phenotype, through increasing the number of perinuclear lysosomes while removing lysosomes form cellular protrusions, was evaluated for each screened drug, using the ImageXpress Micro Confocal High-Content Imaging System. We have hereby identified eight drugs that can redirect lysosomes from the invadosome-like cellular protrusions to their non-invasive perinuclear position in ErbB2 expressing breast cancer cells. We are currently analyzing the impact of these drugs on breast cancer cell invasion though migration assays and three-dimensional invasion assays. We are also addressing their effect on lysosomal membrane permeabilization, lysosomal pH and investigating their effect on proteins that are known to be involved in lysosomal distribution.

Clinically, our findings may facilitate repurposing or new development of drugs to treat HER2/ErbB2-positive and invasive breast cancers that are resistant to current treatments.

Citation Format: Malene B. Hansen, Maria Postol, Davig A. Egan, Marja Jäättelä, Tuula Kallunki. Inhibition of invasion of HER2-positive breast cancer cells by lysosome targeting drugs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4593.
Abstract 4601: Phospholipid and enzyme antibodies for the evaluation of novel cancer biomarkers
Authors: Day Cameron , Neilsen Paul
Published: August 2019

Sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) are well characterized bioactive phospholipids with migratory and proliferative effects in cancer progression. Recent studies show that the enzyme responsible for production of S1P, sphingosine kinase (SK), is upregulated in a variety of cancers, S1P is a tumorigenic growth factor, and there is an association between S1P levels in cancer patients and their resulting clinical outcomes. Similarly, LPA has been shown to drive angiogenesis and tumor invasion. LPA receptors and the LPA synthesizing enzyme autotaxin (ATX) have also been shown to be abnormally expressed in at least ovarian and breast tumors. S1P, SK, LPA, and ATX therefore represent potential biomarkers for the detection and staging of cancer. We have now characterized anti-S1P and anti-LPA antibodies for basic immunochemical methods in cells and ovary tissue. These antibodies were previously developed as anti-cancer immunotherapeutic agents. Here we highlight the potential for these reagents as bonafide cancer biomarkers for S1P and LPA.

Citation Format: Cameron Day, Paul Neilsen. Phospholipid and enzyme antibodies for the evaluation of novel cancer biomarkers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4601.
Abstract 4625: Mouse strains for cancer research at The Jackson Laboratory Repository
Authors: Boswell Deborah , Rockwood Stephen , Lutz Cathleen ,   The JAX Repository Team
Published: August 2019

Genetically defined mice have been the cornerstone of cancer research and resources at The Jackson Laboratory since the founding of the laboratory nearly nine decades ago. The Jackson Laboratory distributes more than 10,500 strains to the scientific community, many with applications in cancer research, including mouse models for specific cancers, xenograft models, immunodeficient platforms for PDX studies, multi-purpose “tool strains” (such as CRISPR cas9-expressing lines), recombinase expressing strains, as well as conditional and inducible expression lines (such as Cre-lox). New to the JAX Repository collection is a KrasLSL-A146T strain with inducible mutant Kras A146T for use in research of gastrointestinal tract and hematological cancers. The tamoxifen-inducible model for pancreatic ductal adenocarcinoma (PDAC), KPCX, and the related conditional activatable Kras allele/floxed Trp53 double mutant line, KP, have recently been developed. Inducible expression of mutant human speckle-type POZ protein (SPOP) in one newly acquired line allows study of prostate tumorigenesis. The portfolio of strains useful for engraftment studies has expanded, complementing the versatile humanized NOD scid gamma (NSG) platform lines. For custom generation of mutant strains, the Repository distributes a variety of research tool strains such as cas9-expressing lines, both constitutive expressing and Cre-inducible models, on different defined genetic backgrounds. Other recently added mutants allow research of the role of inflammation in cancer and screening immunomodulatory cancer therapies.

In addition to safeguarding each strain by cryopreservation, the Repository‘s comprehensive quality control program confirms mutation identity and genetic background, and screens for unwanted alleles (such as stray GFP, cre, lacZ, etc.). Investigators can search for specific strains on the JAX Mice website, or peruse the Oncology Therapeutic Area webpage for cancer research related strains and resources. Researchers can submit their strains to be considered for inclusion in the Repository on The Jackson Laboratory website:

www.jax.org/donate-a-mouse

The Jackson Laboratory Repository is supported by the NIH, The Howard Hughes Medical Institute, and other private charitable foundations.

Citation Format: Deborah Boswell, Stephen Rockwood, Cathleen Lutz, The JAX Repository Team. Mouse strains for cancer research at The Jackson Laboratory Repository [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4625.
Abstract 4635: Newly established mouse oral carcinoma cell lines and their syngeneic tumorigenesis models
Authors: Chen Yi-Fen , Liu Chung-Ji , Kao Shou-Yen , Lin Shu-Chun , Chang Kuo-Wei
Published: August 2019

Oral squamous cell carcinoma (OSCC) is one of the most prevalent malignancies worldwide. Despite the advances in diagnosis and treatment, the survival of OSCC remains to be improved. miR-211 is an oncogenic microRNA frequently disrupted in human OSCC and its high expression is a determinant of patient’s poor survival. Herein, we established four murine OSCC cell lines, designated MOC-L1 - MOC-L4 from the tongue tumor tissues induced by 4-nitroquinoline 1-oxide in K14-EGFP-miR-211 transgenic mice. All cell lines appear green fluorescence and express epithelial markers. The gene expression profiles among cell lines are complex and diverse, while MOC-L1 - MOC-L3 cells carry missense mutations in p53 gene. MOC-L1 exhibits tremendous epithelial-mesenchymal transition and the associated aggressive characteristics. On the contrary, MOC-L4 displays the least invasiveness. Both MOC-L1 and MOC-L2 are clonogenic in vitro and tumorigenic when implemented into dermis or tongue in syngeneic recipients. But, only MOC-L1 exhibits high potential for local regional and distal metastasis. Since the expression of miR-196b in MOC-L1 xenografts drastically decreased upon cisplatin treatment, targeting of miR-196b might facilitate tumor abrogation. As these cell lines originate from the C57BL/6 mouse, which is the strain most suitable for transgenic engineering, to approach the interplay of these OSCC cells with other type of genetically modified cells in immune-competent mice would bestow profound insight on OSCC pathogenesis.

Note: This abstract was not presented at the meeting.

Citation Format: Yi-Fen Chen, Chung-Ji Liu, Shou-Yen Kao, Shu-Chun Lin, Kuo-Wei Chang. Newly established mouse oral carcinoma cell lines and their syngeneic tumorigenesis models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4635.
Abstract 4650: TOPK is a prognostic marker and therapeutic target in skin carcinogenesis
Authors: Roh Eunmiri
Published: August 2019

Solar ultraviolet (sUV) is linked to development of cutaneous squamous cell carcinoma (cSCC) as a non-melanoma skin cancer (NMSC). Actinic keratosis (AK), induced by sUV irradiation, acts as precursor to cutaneous squamous cell carcinoma (cSCC) and is a pre-malignant skin lesion characterized with an abnormal proliferation of atypical keratinocytes confined to the epidermis of the skin. Therefore, to identify the major signaling molecules in the prevention, development and metastasis of cSCC is a critically important task. Here, we suggested a promising protein target, T-LAK cell-originated protein kinase (TOPK) as an active form of MEK1 continuously phosphorylating downstream targets, which appears to be critical in mediating skin carcinogenesis. We detected high levels of total and phosphorylated TOPK protein expression in SCC, and phosphorylated TOPK levels were also markedly higher in AKs and SCCs from patients compared to matched normal skin. Inflammation is known to contribute to tumor development and thus we examined the effect of a 2-minimal erythema dose (MED) of solar stimulated light (SSL) exposure in clinical human skin samples. Our results indicated that acute SSL irradiation increased epidermal thickness as a feature of inflammation and also enhanced total protein and phosphorylation levels of TOPK in human skin in a time-dependent manner. TOPK phosphorylation in human precancerous HaCaT keratinocytes was also increased by SSL irradiation in a dose- and time-dependent manner. Interestingly, TOPK knockout (TOPK-/-) SKH1 (Crl: SKH1-Hrhr) hairless mice showed no tumors on SSL-exposed skin tissues. In contrast, wild-type (TOPK+/+) control SKH1 hairless mice treated with SSL showed increased tumor volume and number. Furthermore, TOPK-/- mice showed base levels of epidermal thickness similar to the SSL-unexposed control group and the Ki-67 proliferation marker was decreased in SSL-exposed TOPK-/- mouse skin compared to SKH1 TOPK+/+ mice. Notably, levels of the anti-apoptotic protein Bcl-2 were decreased in TOPK-/- mice, compared to SSL-treated TOPK+/+ mice. Moreover, our results indicated that phosphorylated and total TOPK protein levels in SCC metastasized to skin or lymph nodes were significantly higher compared to human SCC primary tumors. These results indicated that phosphorylated and total TOPK proteins are associated with both SCC development and metastasis. Thus, molecular targeting of TOPK could be an effective approach in the clinic to treat or prevent cSCC.

Citation Format: Eunmiri Roh. TOPK is a prognostic marker and therapeutic target in skin carcinogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4650.
Abstract 4658: Cigarette smoke extract and benzo(a)pyrene induce osteopontin expression in lung cancer and promote mesenchymal stem cell recruitment
Authors: Jiang Ya-Jing , Chang An-Chen , Chen Po-Chun , Tang Chih-Hsin
Published: August 2019

Cigarette smoking produces many organic compounds, including the carcinogenic polycyclic aromatic hydrocarbon benzo(a)pyrene, (BAP), which is linked to lung cancer. Tumor tissue recruits mesenchymal stem cells (MSCs; undifferentiated multipotent adult stem cells) to increase cancer cell proliferation, invasion and metastasis. We sought to determine the effects of cigarette smoke extract (CSE) and BAP stimulation on lung cancer cells and MSCs. We found that CSE and BAP increased mRNA and protein expression of osteopontin (OPN), a member of the small integrin-binding ligand N-linked glycoproteins (SIBLINGs) family, but had no such effects on any other SIBLINGs proteins. A National Center for Biotechnology Information (NCBI) analysis of data from the Gene Expression Omnibus (GEO) database (GSE31210) identified much higher OPN expression in current and ex-cigarette smokers with lung cancer than in lung cancer patients who were nonsmokers. Our analysis found that stimulating lung cancer cells with CSE and BAP led to the recruitment of MSCs, which adhered to lung cancer cells via OPN production. Next, we used a gene set from the BioCarta database to investigate signaling pathways associated with lung cancer. We found that the MAPK, JAK/STAT and RAS signaling pathways had the highest repeatability in lung cancer patients who were cigarette smokers. We discovered that CSE and BAP only activated the JAK2/STAT3 pathway and that transfection of lung cancer cells with JAK2 and STAT3 small interfering RNAs interrupted CSE- and BAP-induced OPN expression, inhibiting MSC recruitment and adhesion. We are now investigating whether MSCs link back to lung cancer cells and further induce migration and invasion. In summary, our data indicate that CSE and BAP increase OPN expression in lung cancer cells through the JAK2/STAT3 signaling pathway and thereby recruit MSCs and their adhesion to cancer cells.

Citation Format: Ya-Jing Jiang, An-Chen Chang, Po-Chun Chen, Chih-Hsin Tang. Cigarette smoke extract and benzo(a)pyrene induce osteopontin expression in lung cancer and promote mesenchymal stem cell recruitment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4658.
Abstract 4859: MFP-FePt-GO nanocomposite promotes radiosensitivity of non-small cell lung cancer via activating caspase system and impairing DNA damage repair
Authors: Peng Shan , Sun Yingming , Luo Yuan , Ma Shijing , Gong Yan , Xie Conghua
Published: August 2019

Recent advances in nanomedicine provide promising alternatives for cancer treatment that may improve the survival and life quality of patients with cancer. A novel nanocomposite (MFP-FePt-GO) based on a lamellar-structure magnetic graphene oxide (GO) and polyethylene glycol drug delivery system was designed, synthesized and functionalized for co-delivery of metronidazole and 5-fluorouracil. The chemical synthesis efficiency was measured by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy and energy dispersive X-ray spectroscope. The morphology, dispersion and sizes of this nanocomposite were also measured with transmission electron microscope. This new nanocomposite exhibited good solubility, stability and biocompatibility. While no severe allergies, liver and kidney damage or drug-related deaths were observed, MFP-FePt-GO promoted radiosensitivity of non-small cell lung cancer (NSCLC) cells both in vivo and in vitro. MFP-FePt-GO imrpvoed the effects of radiation through activation of the caspase system and impairment of DNA damage repair. This novel nanocomposite also induced a reactive oxygen species burst, which suppressed the antioxidant protein expression and induced cell apoptosis. Furthermore, MFP-FePt-GO prevented the migration and invasion of NSCLC cells. Taken together, MFP-FePt-GO showed a synergistic anti-tumor effect with radiation in eliminating the tumors. With good safety and efficacy, this novel nanocomposite might be a potential radiosensitive agent for NSCLC patients.

Citation Format: Shan Peng, Yingming Sun, Yuan Luo, Shijing Ma, Yan Gong, Conghua Xie. MFP-FePt-GO nanocomposite promotes radiosensitivity of non-small cell lung cancer via activating caspase system and impairing DNA damage repair [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4859.
Abstract 487: A high percentage of LCNEC shows DLL3 expression in association with molecular subtypes and neuroendocrine markers
Authors: Speel Ernst-Jan M. , Hermans Bregtje C. , Derks Jules L. , Thunnissen Erik , Suylen Robert Jan van , Bakker Michael A. den , Groen Harry J. , Smit Egbert F. , Damhuis Ronald A. , Broek Esther C. van den , Stallinga Cecile M. , Roemen Guido M. , Dingemans Anne-Marie C.
Published: August 2019

Background Pulmonary large cell neuroendocrine carcinoma (LCNEC) can be subdivided in two types: the co-mutated TP53 and RB1 subtype, and the TP53 and STK11/KEAP1 mutated subtype (Derks et al. Clin Cancer Res 2018; J Thorac Oncol 2018). DLL3 is a member of the Notch ligand family and a possible treatment target for neuroendocrine carcinoma. We investigated DLL3 and NE marker expression in mutational subtypes of metastatic LCNEC.

Methods Immunohistochemical (IHC) analysis for DLL3 (clone SC16.65) was performed on 94 pathological reviewed pretreatment stage IV LCNEC. Samples were scored positive if ≥1% tumor cells showed cytoplasmic or dotlike DLL3 immunostaining. Also an H-score was calculated by multiplying intensity by percentage of positive cells. Targeted next generation sequencing (TP53, RB1, STK11, KEAP1) could be performed in 66 patients.

Results DLL3 was expressed in 70/94 (75%) LCNEC, 56 of which showed cytoplasmic immunostaining. 37/70 (53%) samples had an H-score>100. DLL3 staining was more often seen in STK11 and/or KEAP1 mutated LCNEC (13/14;93%) than in tumors with wildtype STK11/KEAP1 (36/52;69%) (trend;p=0.093). In addition, DLL3 positivity was associated with expression of ≥2 NE-markers (64/79 LCNEC (81%) vs 3/8 (37%) tumors with <2 NE-markers; p=0.014).

Conclusions A high percentage (75%) of stage IV LCNEC shows cytoplasmatic DLL3 epression in association with NE markers, half of which with H-scores>100. Interestingly, DLL3 positivity was observed in almost all STK11/KEAP1 mutated LCNEC, which is in agreement with recent data (George et al. Nat Commun 2018). It thus might be worthwhile to investigate the efficacy of DLL3 targeted therapy in LCNEC.

Citation Format: Ernst-Jan M. Speel, Bregtje C. Hermans, Jules L. Derks, Erik Thunnissen, Robert Jan van Suylen, Michael A. den Bakker, Harry J. Groen, Egbert F. Smit, Ronald A. Damhuis, Esther C. van den Broek, Cecile M. Stallinga, Guido M. Roemen, Anne-Marie C. Dingemans. A high percentage of LCNEC shows DLL3 expression in association with molecular subtypes and neuroendocrine markers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 487.
Abstract 4870: Dedicator of cytokinesis-x promotes chemo and radioresistance of gastric cancer by modulating WNT signaling and cancer stem cell traits
Authors: Chi Hsiang-Cheng , Lin Kwang-Huei
Published: August 2019

Prognosis of gastric cancer (GC) remains poor and the key players in its molecular pathogenesis are unknown at present. There is therefore a critical need to identify novel prognostic factors of GC. Dedicator of cytokinesis represents a protein family belonging to atypical Rho guanine nucleotide exchange factors (GEF) for Rac and/or Cdc42 GTPases. Dedicator of cytokinesis-x, a member of the Dock-C subfamily, exchanges GDP for GTP for Rac1 and Cdc42, both in vitro and in vivo. Here, we identified Dedicator of cytokinesis-x as an independent biomarker for GC prognosis. Elevated Dedicator of cytokinesis-x expression was associated with significantly shorter cumulative survival in both univariate and multivariate analyses. Moreover, higher dedicator of cytokinesis-x expression is positively correlated with tumor size, depth of invasion, lymph node metastasis, vascular invasion and pathological stage. To establish the downstream effectors of dedicator of cytokinesis-x, we analyzed the genes in relation to dedicator of cytokinesis-x expression using three independent clinical GC cohorts. Genes displaying a positive correlation with dedicator of cytokinesis-x were subsequently subject to gene ontology analysis and shown to be significantly involved in the WNT/β-catenin signaling pathway. Depletion of dedicator of cytokinesis-x suppressed cell proliferation, chemo- and radio-resistance, metastasis, oncosphere formation ability and tumorigenicity as well as expression of gastric cancer stem cell (CSC) markers. Dedicator of cytokinesis-x depletion was accompanied by a concomitant decrease in CD44, CD133 and nuclear β-catenin expressions. Moreover, interactions between dedicator of cytokinesis-x and β-catenin were observed in the nucleus of GC cells. This finding suggested that dedicator of cytokinesis-x may act as a tumor promoter through WNT/β-catenin activation.

Citation Format: Hsiang-Cheng Chi, Kwang-Huei Lin. Dedicator of cytokinesis-x promotes chemo and radioresistance of gastric cancer by modulating WNT signaling and cancer stem cell traits [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4870.
Abstract 4887: Recurrent NRG1 rearrangements in Caucasian pulmonary mucinous adenocarcinoma: results from an Italian multi-center cohort
Authors: Trombetta Domenico , Graziano Paolo , Sparaneo Angelo , Rossi Giulio , Rossi Antonio , Tiseo Marcello , Maio Massimo Di , Fabrizio Federico P. , Manzorra Maria C. , Centra Flavia , Candia Leonarda Di , Audisio Marco , Maiello Evaristo , Fazio Vito M. , Muscarella Lucia A.
Published: August 2019

Invasive Mucinous Adenocarcinoma (IMA) is a rare histotype of lung adenocarcinoma associated with an unfavorable prognosis due to the lack of effective treatment. The NRG1 rearrangement is a new subtype-specific molecular feature of IMA and acts as a strong oncogenic inductor of the aberrant tyrosine kinase activity of ErbB2/ErbB3 heterodimers through PI3K-AKT/MAPK cellular cascades. We recently described for the first time the occurrence of NRG1 rearrangements in 31% of Caucasian lung IMAs and highlighted a strong association between NRG1 rearrangements and ErbB3 activation.

Here we extended our lung IMA samples cohort by enrolling a total of 71 patients from three different Italian Centers and collected clinical-pathological information and molecular profile, included the mutational status of KRAS, EGFR and ALK genes. We screened all samples by fluorescent in situ hybridization (FISH) for the detection of putative NRG1 rearrangements and by immunohistochemistry (IHC) for the expression of phosphorylated-ErbB3 (pErbB3) receptor. Finally, we customized a new targeted RNA Custom Panel to detect all 9 NRG1-fusion variants published to date to molecular characterize the NRG1 fusion variants in NRG1 rearranged IMAs.

Results showed NRG1 rearrangements in 32% of lung IMAs, displaying both NRG1 FISH split signals and deletions of the 5' portion of the gene. IHC confirmed our previous findings of association between pErbB3 immunoreactivity and NRG1 rearrangements, and the heterogeneity of fluorescent signal distribution and immunostaining along the tissue sections. The CD74-NRG1 remains the most common fusion variant identified in lung IMA samples. Correlation analysis among clinical-pathological data, pErbB3 expression and NRG1 rearrangements are ongoing.

Our results confirm the usefulness of IHC/FISH combined approach for NRG1 broken tumors identification and highlight the role of NRG1 rearrangement as master molecular marker of lung IMAs, potentially useful to select patients for the emerging target therapies.

Citation Format: Domenico Trombetta, Paolo Graziano, Angelo Sparaneo, Giulio Rossi, Antonio Rossi, Marcello Tiseo, Massimo Di Maio, Federico P. Fabrizio, Maria C. Manzorra, Flavia Centra, Leonarda Di Candia, Marco Audisio, Evaristo Maiello, Vito M. Fazio, Lucia A. Muscarella. Recurrent NRG1 rearrangements in Caucasian pulmonary mucinous adenocarcinoma: results from an Italian multi-center cohort [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4887.
Abstract 4895: Natural history and outcome of patients presenting a metastatic breast cancer with PIK3CA mutation
Authors: Mosele Fernanda , Verret Benjamin , Lusque Amelie , Filleron Thomas , Bachelot Thomas , Arnedos Monica , Campone Mario , Dalenc Florence , Lefeuvre Claudia , Sablin Marie Paule , Bonnefoi Hervé , Lacroix Ludovic , Bièche Ivan , Gonçalves Anthony , Jacot William , Jimenez Marta , Jacquet Amelie , Andre Fabrice , Andre Fabrice
Published: August 2019

BACKGROUND: Activating PIK3CA mutations occur in 20-30% of patients with metastatic breast cancer (mBC). A recent study showed that alpha selective PI3K inhibitors improve outcome in patients with PIK3CA mutation, HR+/Her2- mBC. There is a need to better understand the natural history of PIK3CA mutant mBC to optimally position of this drug family.

PATIENTS AND METHODS: 649 patients from SAFIR02 trial (NCT02299999), for which mutational profile was available and with clinical data entered in a database, were selected for this analysis. Associations between PIK3CA mutation, clinical characteristics and outcome were analyzed.

RESULTS: 143 patients (22%) harbor PIK3CA mutation in tumor sample. 10% (n=27) and 34% (n=104) of TNBC and HR+/Her2- mBC presented a PIK3CA mutation respectively (p<0.001). In patients with HR+/Her2- mBC, there is no significant association between PIK3CA mutation and site of metastases or number of metastases (p<0.01). In the same group, patients with PIK3CA mutations were less sensitive to chemotherapy (stable disease or response: 51.3% vs. 69.2% in PIK3CA wild type; p=0.005) (Odds ratio multivariate: 0.40 [95% CI: 0.22-0.71] p=0.002). The median overall survival (OS) for patients with PIK3CA mutated HR+/Her2- mBC was of 19.6 months vs. 23.5 for patients with PIK3CA wild type (p=0.048) (HR multivariate: 1.44 [95% CI: 1.02-2.03] p=0.039). HR+/Her2- mBC with PIK3CA mutations presented more frequently a MAP3K1 mutation (16% vs. 4%, p=0.0002). In the TNBC group, the median OS in patients with PIK3CA mutated cancer was of 24.2 months vs. 14 months in the wild type group (p=0.028).

CONCLUSION: Patients with PIK3CA mutated, HR+/Her2- mBC are less sensitive to chemotherapy and present a shorter survival. These data emphasize the need for new therapies in this setting, and to position these therapies earlier than chemotherapy in the treatment sequences.

Citation Format: Fernanda Mosele, Benjamin Verret, Amelie Lusque, Thomas Filleron, Thomas Bachelot, Monica Arnedos, Mario Campone, Florence Dalenc, Claudia Lefeuvre, Marie Paule Sablin, Hervé Bonnefoi, Ludovic Lacroix, Ivan Bièche, Anthony Gonçalves, William Jacot, Marta Jimenez, Amelie Jacquet, Fabrice Andre, Fabrice Andre. Natural history and outcome of patients presenting a metastatic breast cancer with PIK3CA mutation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4895.
Abstract 49: ET-1 receptor blockade in engineered 3D high-grade serous ovarian cancer tumoroids
Authors: Rosanò Laura , Pape Judith , Cheema Umber , Loizidou Marilena , Bagnato Anna
Published: August 2019

High-grade serous ovarian cancer (HG-SOC), accounting for 70-80% of ovarian cancer deaths, is characterized by widespred and rapid metastatic nature, suggesting that unraveling determinants of metastasis may implement new therapeutic strategies on clinical care. In this tumor, autocrine or paracrine activation of the endothelin-1 receptors (ET-1R) are recognized drivers of tumor progression, promoting epithelial-mesenchymal transition (EMT), invasion, and metastasis, as well as tumor angiogenesis and lymphangiogenesis. However, since the metastatic spread and response to therapy are influenced by the extracellular matrix (ECM) density and composition of the surrounding tumour microenvironment (TME), the development of a three-dimensional in vitro model mimicking the in vivo tumors is necessary to better understand the dynamic interactions between tumor cells and TME elements, including fibroblasts, endothelial cells and the ECM, regulated by ET-1R. With this in mind, we have developed HG-SOC tumoroids by engineering a dense central artificial cancer mass (ACM) containing HG-SOC cells, nested within a compressed hydrogel representing the stromal compartment comprising type I collagen, laminin, fibronectin, and stromal cells (fibroblasts and endothelial cells). After 21 days, ET-1-stimulated HG-SOC cells showed an altered migration pattern and formed cellular aggregates, mimicking micrometastases that invaded the stroma. As compared to control cells, ET-1 treated cells showed a higher number of cellular aggregates, which were reduced by treatment with the dual receptor antagonist macitentan. In addition, ET-1 increased the size of the invading aggregates compared to control cells. These findings suggest that HG-SOC tumoroids might be useful to investigate the cross-talk between HG-SOC cells and TME in cancer progression and the response to combinatorial treatment, including ET-1R antagonist.

Note: This abstract was not presented at the meeting.

Citation Format: Laura Rosanò, Judith Pape, Umber Cheema, Marilena Loizidou, Anna Bagnato. ET-1 receptor blockade in engineered 3D high-grade serous ovarian cancer tumoroids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 49.
Abstract 4904: High levels of microRNA-210-3p are associated with increased risk of disease progression in breast cancer patients treated with docetaxel
Authors: PASCULLI BARBARA , BARBANO RAFFAELA , RENDINA MICHELINA , FONTANA ANDREA , COPETTI MASSIMILIANO , VALORI VANNA MARIA , MORRITTI MARIA , MAIELLO EVARISTO , FAZIO VITO MICHELE , ESTELLER MANEL , PARRELLA PAOLA
Published: August 2019

MiR-210-3p is the most hypoxia-sensitive miRNA showing overexpression in different human cancers, but its role as biomarker in breast cancer (BC) is still controversial. In our study, we performed the expression analysis of miR-210-3p in a retrospective cohort of breast cancer patients with a median follow-up of 91 months enrolled according to the REMARK guidelines (n=284). As controls, 13 normal breast tissues (NBTs) from reductive mammoplasty were also profiled. By this analysis, we found that miR-210-3p levels were significantly increased in tumours as compared with NBTs (P=0.021). According to the surrogate molecular classification, the Triple Negative subgroup showed the highest levels of miR-210, followed by Luminal B tumours, whereas Luminal A, and HER2-amplified tumours showed the lowest expression levels (P=0.0137). The evaluation of the association of miR-210-3p with time-to-event outcomes in patients without synchronous metastases (n=268) demonstrated an association between risk of disease progression (HR 2.13, 95%CI 0.33-3.39, P=0.002) and higher levels of miR-210-3p in the subgroup treated with docetaxel in sequential therapy with anthracyclines (ECD). Moreover, a cut-off value of 20.966 as established by ROC curve analysis allowed to discriminate patients who developed distant metastases with an accuracy of 85% at 3-years (AUC 0.87, CI 0.69-1) and 83% at 5-years follow up (AUC 0.832, CI 0.656-1). Instead, the accuracy in discriminating patients who died for the disease was of 79.6% at both 5- (AUC 0.804, CI 0.517-1) and 10-years (AUC 0.804 CI 0.517-1) follow up. Overall, our data support the putative involvement of miR-210-3p in the response to treatments including docetaxel and suggest its eligibility to become a biomarker of early metastases development in ECD-treated breast cancer patients.

Citation Format: BARBARA PASCULLI, RAFFAELA BARBANO, MICHELINA RENDINA, ANDREA FONTANA, MASSIMILIANO COPETTI, VANNA MARIA VALORI, MARIA MORRITTI, EVARISTO MAIELLO, VITO MICHELE FAZIO, MANEL ESTELLER, PAOLA PARRELLA. High levels of microRNA-210-3p are associated with increased risk of disease progression in breast cancer patients treated with docetaxel [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4904.