bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1569: TGFβ signaling suppresses alternative end joining associated with genomic mutations and PARP Inhibition sensitivity
Authors: Liu Qi , Palomero Luis , Moore Jade , Guix Ines , Pujana Miquel Angel , Barcellos-Hoff Mary Helen 
Published: September 2020

Abstract
Differential use of DNA repair pathways in cancer leads to distinct clinical outcomes following genotoxic therapies. We found that deficient TGFβ signaling due to human papilloma virus (HPV) in head and neck squamous cell carcinoma (HNSCC) results in impaired DNA repair proficiency, increased use of alternative end joining (alt-EJ) and that a 50-gene TGFβ signature is negatively correlated with prognosis (Clin Ca Res 2018, 24(23): 6001). To assess the molecular consequence of TGFβ signaling in the DNA damage response (DDR), we performed a Nanostring assay on SAS HNSCC and U251 glioblastoma cell lines using the 180 gene DNA damage and repair panel. The results showed that treatment with TGFβ type 1 receptor kinase inhibitor LY2157299 or TGFβ profoundly altered DDR gene expression profiles. TGFβ inhibition for 24 hr significantly (p&lt;0.05) increased critical alt-EJ genes, POLQ (DNA polymerase theta) and PARP1 (poly(ADP-ribose) polymerase 1), which was confirmed by qRT-PCR. To interrogate the relationship between TGFβ and alt-EJ in The Cancer Genome Atlas (TCGA), we used the literature to curate a 36 gene alt-EJ signature. Single specimen gene set enrichment analyses (ssGSEA) showed that the alt-EJ signature was significantly anti-correlated with the TGFβ signature in 15 of 17 cancer types (PCC = -0.35; p =10-16). This anti-correlation was also observed in 875 cancer cell lines in Catalogue of Somatic Mutations in Cancer (COSMIC) database, suggesting a conserved, cancer cell intrinsic mechanism by which TGFβ regulates alt-EJ. Alt-EJ is an error prone DNA repair mechanism that generates gross genomic alterations. Consistent with this, low ssGSEA score for TGFβ and high ssGSEA score for alt-EJ are correlated with increased deletions or insertions in genome sequence of patient tumors in TCGA as well as cancer cell lines in COSMIC (p&lt;0.05). PARP1 requirement prompted analyses of drug sensitivity data in the Sanger database, which showed that cell lines with high alt-EJ and low TGFβ ssGSEA score are significantly more sensitive to PARP1 inhibitors, olaparib and ABT-888, (p &lt; 0.0001). These data lead to the novel hypothesis that TGFβ signaling actively suppresses alt-EJ. If true, the TGFβ/alt-EJ signature may be useful as a biomarker for precision cancer medicine with genotoxic agents, TGFβ or PARP inhibitors.
Citation Format: Qi Liu, Luis Palomero, Jade Moore, Ines Guix, Miquel Angel Pujana, Mary Helen Barcellos-Hoff. TGFβ signaling suppresses alternative end joining associated with genomic mutations and PARP Inhibition sensitivity [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1569.
Abstract 1582: A pre-existing inflammatory immune microenvironment predicts the clinical and immunological response of vulvar high-grade squamous intraepithelial lesions to therapeutic HPV16 peptide vaccination
Authors: Abdulrahman Ziena , de Miranda Noel F. , van Poelgeest Mariette I. , van der Burg Sjoerd H. 
Published: September 2020

Abstract
Background: Vulvar High-grade Squamous Intraepithelial Lesion (vHSIL) is predominantly induced by high-risk Human Papilloma Virus type 16 (HPV16). In two independent trials, therapeutic vaccination against the oncoproteins of HPV16 with synthetic long peptides (SLP) resulted in vHSIL regression in about half of the patients after 12 months. Several studies have shown that the immune microenvironment influences therapy outcome. Therefore, a thorough investigation of the vHSIL immune microenvironment before and after SLP vaccination was performed, and its impact on clinical response was studied.
Methods: Two novel multiplex immunofluorescence panels were designed and fully optimized for formalin-fixed paraffin-embedded tissue, one for T cells (CD3, CD8, Foxp3, Tim3, Tbet, PD-1, DAPI) and one for myeloid cells (CD14, CD33, CD68, CD163, CD11c, PD-L1, DAPI). Pre- and post-vaccination biopsies of 29 vHSIL patients, as well as 27 healthy vulva excisions, were stained, scanned with the Vectra multispectral imaging system, and automatically phenotyped and counted with inForm advanced image analysis software.
Results: A pre-existing pro-inflammatory tumor microenvironment, marked by high numbers of CD4+ and CD8+ T cells expressing Tbet and/or PD-1 as well as CD14+ inflammatory macrophages, is a strong predictor for good clinical response to therapeutic HPV16 SLP vaccination. A clear stepwise increase in pre-vaccination infiltrating Tbet+, CD4+, CD8+ T cells and CD14+ macrophages, and decrease in Foxp3+ regulatory T cells was observed as response increased from non to partial to complete response. Moreover, the pre-vaccination immune microenvironment of complete responders resembled healthy vulva. Vaccination further increased infiltrating CD4+ and Tbet+ T cells and CD14+ macrophages, and decreased Foxp3+ regulatory T cells and CD68+CD163+ M2 macrophages in the complete and partial responders, but not in the non responders.
Conclusion: Clinical responsiveness to therapeutic HPV16 SLP vaccination requires a pre-existing inflamed type 1 immune contexture in vHSIL. Hence, only patients with an inflamed microenvironment should be selected for monotherapy by therapeutic peptide vaccination, since this strategy is incapable of overcoming an immunologically cold tumor microenvironment.
Citation Format: Ziena Abdulrahman, Noel F. de Miranda, Mariette I. van Poelgeest, Sjoerd H. van der Burg. A pre-existing inflammatory immune microenvironment predicts the clinical and immunological response of vulvar high-grade squamous intraepithelial lesions to therapeutic HPV16 peptide vaccination [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1582.
Abstract 1608: Adaptive resistance to PD-1 blockade upregulates Tim-3 through STAT3 pathway in regulatory T cells in melanoma
Authors: Huang Lili , Xu Yu , Fang Juemin , Liu Weixing , Liu Zhuqing , Xu Qing 
Published: September 2020

Abstract
Tregs play an opposing role in anti-tumour immunity by suppressing effective immunotherapies in melanoma. Thus, understanding the precise mechanisms of Tregs in immune therapy is important. TILs separated from freshly excised melanoma tumours and PBMCs obtained from healthy donors were used to assess the expression levels of Tim-3 and IL-10/TGF-β of Tregs by multi-colour flow cytometry and immunohistochemistry in the presence or absence of an anti-PD-1 antibody or STAT3 inhibitor in vitro and in vivo. The mRNA level of STAT3 gene was closely associated with the expression of PD-1 and Tim-3 in melanoma patients. Tim-3 expression was significantly upregulated on Tregs after PD-1 blockade in mice treated with either induced mouse and human melanoma TILs or healthy donor PBMCs. The levels of IL-10 and TGF-β secretion in the TIL group increased compared with those in the control group, supporting a potential escape mechanism from anti-PD-1 therapy in the tumour microenvironment. Furthermore, in the presence of the STAT3 pathway inhibitor Stattic, Tim-3 and PD-1 expression on Treg was regulated on TILs from respondent mice melanoma, suggesting that STAT3 could mediate Tim-3 and PD-1 expression on Treg in tumor microenvironment. Therefore, our findings reveal the mechanism of PD-1 and Tim-3 expression in a STAT3-dependent manner on Tregs in the TME, providing further support for STAT3 as a potential transient small molecular inhibitor targets for patients suffering from melanoma.
Citation Format: Lili Huang, Yu Xu, Juemin Fang, Weixing Liu, Zhuqing Liu, Qing Xu. Adaptive resistance to PD-1 blockade upregulates Tim-3 through STAT3 pathway in regulatory T cells in melanoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1608.
Abstract 1611: MYB and the MYB-NFIB fusion induce adenoid cystic carcinoma
Authors: Hu Yuan , Zhu Xiaoyuan , Luo Han , El-Naggar Adel , Caulin Carlos 
Published: September 2020

Abstract
Adenoid cystic carcinoma (ACC) is the second most common malignancy of the salivary glands, also found at lower frequency rates in other secretory glands such as the mammary and lacrimal glands. Salivary ACCs are slow-growing but aggressive neoplasms, with propensity to perineural invasion and high recurrence rates. MYB is the most frequently altered gene in ACCs, as approximately 40% of the ACCs contain MYB-NFIB gene fusions, and the full-length MYB is overexpressed in over 50% of the fusion-negative ACCs. However, it is still unclear whether MYB activities promote ACC development. To assess the oncogenic potential of MYB in vivo, we generated transgenic mice that overexpress MYB using the TetO system. In these mice, focal MYB overexpression was directed to salivary and mammary glands using the activator line MMTV-tTA. We found that 25% of these mice developed tumors with an average latency of 23 months. Remarkably, all the tumors that developed in these mice were ACCs, and the three main phenotypic variants of human ACCs (cribriform, tubular and solid) were represented in this cohort. Using the same transgenic system, we found that the MYB-NFIB fusion also induced ACCs, with similar incidence and lower latency. Immunohistochemical analysis of these mouse ACCs for several biomarkers of human ACC, including p63 and c-kit, provided additional evidence of the similarity of the mouse ACCs that developed in these models, and human ACCs. RNAseq global expression profiling uncovered the activation of several cancer-associated pathways in MYB-induced mouse ACCs. Most notably, activation NOTCH signaling was highly enriched in these tumors. Additional analysis for the presence of the NOTCH intracellular domain demonstrated that all of the MYB- and MYB-NFIB-induced ACCs contain NOTCH activation. These findings, and the recent identification of activating mutations in NOTCH1 and NOTCH2 in human ACCs, suggest that NOTCH activities may contribute to ACC development. To test this hypothesis, ongoing studies in the lab are directed to assess the role of NOTCH signaling in ACC development, by generating mouse models in which constitutive activation of NOTCH is induced in mice alone or in combination with MYB alterations. Overall, these studies demonstrate that MYB activities are the main oncogenic driver of ACC. Genomic rearrangements that result in MYB-NFIB fusions may enhance the oncogenic potential of MYB.
Citation Format: Yuan Hu, Xiaoyuan Zhu, Han Luo, Adel El-Naggar, Carlos Caulin. MYB and the MYB-NFIB fusion induce adenoid cystic carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1611.
Abstract 1624: Lrig1 is an Egfr-dependent tumor suppressor in mouse duodenal and colonic neoplasia
Authors: Coffey Robert J. , Niitsu Hiro , Lu Yuanyuan , Huh Won Jae , Franklin Jeffrey L. 
Published: September 2020

Abstract
Leucine-rich repeats and immunoglobulin-like domain 1 (Lrig1) is a pan-ErbB negative regulator. Lrig1 null mice develop spontaneous duodenal adenomas by 3-4 months of age but they exhibit no overt colonic pathology (AE Powell Cell 149:146-158, 2012). To examine a possible tumor suppressor role for Lrig1 in colonic neoplasia, we utilized Lrig1CreERT2 and Lrig1Apple mice in which a tamoxifen-inducible Cre recombinase or an Apple fluorescent reporter were inserted into the translational start site of endogenous Lrig1, thus creating Lrig1 null mice. Colonic tumors were generated by administration of AOM/DSS to Lrig1CreERT2/CreERT2 mice and by administration of tamoxifen (TAM) to Lrig1CreERT2/Apple;Apcflox/+ mice to induce loss of one Apc allele. Compared to littermate control mice, there was a significant increase in colonic tumor number in both Lrig1 null cohorts (p=0.016 and p=0.007, respectively).
We previously reported Egfr was the most upregulated ErbB family member in Lrig1 null duodenal tumors (Y Wang Am J Path 185:1123-1134, 2015). To directly implicate Egfr in both Lrig1 null duodenal and colonic tumors, we administered TAM at 6 weeks of age to Lrig1CreERT2/CreERT2;Egfrflox/flox mice. At 6 months of age, all oil-injected mice developed tumors, whereas tumors were not detected in 8/21 (38.1%) of TAM-injected mice; the tumor volume in TAM-injected mice was reduced to 7.78 mm3 (73.5%) compared to (10.58 mm3) in the oil-injected group as measured by MRI scanning (p&lt;0.05). In the AOM/DSS colonic tumor model, administration of TAM at 6 weeks of age to Lrig1CreERT2/CreERT2;Egfrflox/flox mice resulted in a significant decrease in tumor number (p=0.021) but not size compared to oil-injected mice. These results indicate that Lrig1 is an Egfr-dependent tumor suppressor in both mouse duodenal and colonic neoplasia.
Citation Format: Robert J. Coffey, Hiro Niitsu, Yuanyuan Lu, Won Jae Huh, Jeffrey L. Franklin. Lrig1 is an Egfr-dependent tumor suppressor in mouse duodenal and colonic neoplasia [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1624.
Abstract 1626: A novel mouse model of hepatocellular carcinoma induced by c-myc overexpression
Authors: Wang Jinjin , Feng Dan , Zhu Haiyan , Ci Lei , Wan Zhipeng , Sun Ruilin 
Published: September 2020

Abstract
Hepatocellular carcinoma is a fatal liver cancer, and there are limited treatment options for this cancer. c-Myc is a key oncogene in liver cancer, but the specific mechanism of carcinogenesis in liver is unknown. Animal models are widely used to explore the molecular pathogenesis of HCC and test new drug candidates. In this poster, we reported a c-Myc conditional over-expression mouse model (LSL-Myc), which inserts the CAG-Loxp-stop-Loxp-c-Myc-polyA expression cassette into the H11 site. The expression of the Myc gene in this mouse model is regulated by Cre recombinase, which is not normally expressed, but only expressed in Cre-expressing tissue cells. LSL-Myc / Alb-cre double-positive mice were produced by crossing LSL-Myc hybrid mice with Alb-cre mice. In this mouse model, c-Myc was specifically expressed in liver cells. LSL-Myc / Alb-cre double-positive mice show a typical HCC phenotype and show accelerated tumor initiation and rapid HCC progression. Mice can detect significant liver tumorigenesis at 2 months of age, and 100% mice can occur hepatocellular carcinoma. The liver cancer tissues from LSL-Myc / Alb-cre double-positive mice can be transplanted to C57BL/6 mice to establish a mouse-derived PDX model. In summary, this model has the advantages of rapid oncogenesis and high incidence of liver cancer. It is an ideal model for studying the mechanism of c-Myc-driven liver cancer and c-Myc targeted therapy.
Citation Format: Jinjin Wang, Dan Feng, Haiyan Zhu, Lei Ci, Zhipeng Wan, Ruilin Sun. A novel mouse model of hepatocellular carcinoma induced by c-myc overexpression [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1626.
Abstract 1628: Activation of the Nrf2E79Q mutation in mouse lung accelerate development of lung squamous dysplasia
Authors: Hamad Samera H. , Montgomery Stephanie , Bowman Brittany , Murphy Ryan , Major Ben , Weissman Bernard 
Published: September 2020

Abstract
NFE2-related factor 2 (Nrf2) is a transcription factor that regulates more than 500 genes in human cells to protect them from oxidative damage. Somatic mutations in NRF2 resulting in constitutively Nrf2 signaling have been implicated as driver mutations in cancer, especially lung cancer. However, the mechanisms by which NRF2 activating mutations drive tumor development remains unresolved. Therefore, we used a novel genetically engineered mouse model (GEMM), LSL-Nrf2E79Q/+, to understand the role of this activating mutation (one of the most common mutations found in human tumors) in the development of lung tumors. We compared tumor development in two groups of mice: p53fl/fl; p16fl/fl; LSL-Nrf2+/+ (WT, n=38) and p53fl/fl; p16fl/fl; Nrf2E79Q/+ (Het, n=37). We used intranasal instillation of Adenoviral Cre at 6-8 weeks of age to activate the Nrf2E79Q allele and inactivate p53 and p16. Among other tumors, lung tumors were the most common tumors that were found in the GEMM, with major subtypes of lung squamous dysplasia (LSD: pre-malignant lesions of lung squamous cell carcinoma (LSCC)) and lung adenocarcinoma (LADC). Additionally, Het mice showed significantly higher LSD (30%) compared to WT mice (11%) implicating Nrf2E79Q/+ in LSD initiation. However, no significant difference in LADC was observed in Het mice compared to WT mice. Immunohistochemistry (IHC) including Nrf2, TTF1, CK7 and p63 was used to differentiate lung tumors. LADC of WT and Het mice showed negative Nrf2 staining despite the presence of the recombined Nrf2E79Q/+ allele in the Het tumors. However, LSDs developed by Het mice showed positive Nrf2 staining, while those in WT mice were negative, suggesting the mutant Nrf2 increased the frequency of LSDs. LADC of both groups was positive for TTF1 (A primary-non small lung cancer marker, mostly LADC) and CK7 (Cytokeratin7: a marker of epithelial cells). However, LSD lesions were also positive for TTF1. No p63 (a marker of LSCC) stain was shown for LADC of both groups; while p63 was negative to mildly positive in the case of LSD. Results of this study appear consistent with TCGA results where LADC predominantly contained Keap1 loss, while Nrf2 mutations frequently appeared in LSCC. This is the first study to show that a common activating mutation of Nrf2 promotes initiation of LSD that could further develop into LSCC.
Citation Format: Samera H. Hamad, Stephanie Montgomery, Brittany Bowman, Ryan Murphy, Ben Major, Bernard Weissman. Activation of the Nrf2E79Q mutation in mouse lung accelerate development of lung squamous dysplasia [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1628.
Abstract 1659: Development of an orthotopic murine model of a patient-derived pancreatic adenocarcinoma to evaluate the tissue specificity of two novel fluorescent compounds
Authors: Duchamp Olivier , Berthet Cyril , Provent Peggy , Chatel Elodie Marie Dit , Longin Jordan , Urbain Stanislas , Scherninski François 
Published: September 2020

Abstract
Fluorescent dyes have been used in medical diagnostics for over 50 years. The emergence of new fluorescent imaging technology in support of surgery and the need for increased sensitivity and precision have driven the search for novel fluorescent molecules. Orthotopic patient-derived pancreatic tumor models provide a closely related human tumor within organ-specific tumor microenvironment, which was well suited to the evaluation of the tissue specificity of two novel fluorescent compounds.
To this end, we have developed an orthotopic tumor model of IM-PAN-15 (PDX model developed within the IMODI consortium), a patient derived pancreatic adenocarcinoma, in mice. Tumor cells were amplified subcutaneously in SCID mice and the resulting fragments subsequently surgically implanted into the pancreas of irradiated BALB/c nude mice. Tumor development was monitored in vivo by MRI at three time points. Fluorescent compounds were injected intravenously 36 days after tumor fragment implantation, when mean tumor volumes were approximately 70 mm³.
Ex-vivo fluorescent imaging showed that 2h after injection, the two novel fluorescent compounds were present in the pancreas and tumor at roughly equivalent amounts. However, 48h after injection, a clear preferential distribution for the tumor was observed, with both compounds producing an approximately four-fold higher fluorescent signal in the tumor than in the surround pancreatic tissue. This effect persisted six days after injection, although the signal decreased over time. In comparison, indocyanine green, a dye widely used in medical diagnostics, showed no specific accumulation in either the pancreas or tumor.
In conclusion, we report the development of a clinically relevant, patient-derived orthotopic tumor model and demonstrate its use in the investigation of tissue specificity of two novel fluorescent compounds.
Keywords: PDX, fluorescent dyes, orthotopic pancreatic model, MRI
Citation Format: Olivier Duchamp, Cyril Berthet, Peggy Provent, Elodie Marie Dit Chatel, Jordan Longin, Stanislas Urbain, François Scherninski. Development of an orthotopic murine model of a patient-derived pancreatic adenocarcinoma to evaluate the tissue specificity of two novel fluorescent compounds [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1659.