bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1644: Detection and quantification of lymphedema using optical coherence tomography lymphangiography
Authors: Kwan Jennifer Y. , Demidov Valentin , Jones Blake , Shi Wei , Williams Justin , Flueraru Costel , Yip Kenneth W. , Liu Fei-Fei , Vitkin Alex
Published: September 2020

Purpose: Imaging of the lymphatic system is important for monitoring and quantification of the vasculature of tumours and in lymphatic disorders (e.g. lymphedema, an obstruction of the lymphatic system that occurs as a side-effect of cancer treatments such as radiation and/or surgery). Traditional methods of lymphangiography require injection of a contrast agent to visualize the lymphatic system, which may be suboptimal when there is lymphatic obstruction. Thus, there is a need to develop intrinsic-contrast, high-resolution volumetric (3D) non-invasive quantitative in vivo imaging systems for both pre-clinical and potentially clinical use.

Methods: An emerging non-invasive 3D imaging technology called optical coherence tomography (OCT) lymphangiography was optimized to visualize lymphatic vasculature and edema within tissues of C57BL/6 mice under normal and surgically-induced lymphedematous conditions. A murine tail lymphedema model was created by micro-surgical obstruction of the superficial and deep lymphatic networks under a dissecting microscope. OCT imaging was validated against fluorescent microscopy, an established but contrast-agent-dependent lymphangiography technique, as well as histology. Fluorescent microscopy was performed with intradermal injection of FITC-dextran followed by imaging with the AxioZoom.V16 Stereo Zoom Microscope. Immunofluorescence staining was performed with an anti-LYVE1 (Lymphatic Vessel Endothelial Receptor 1) antibody.

Results: Comparing 3D OCT images and their 2D projections under normal and lymphedematous conditions showed significantly different networks. Quantification analysis revealed increased lymphatic vasculature dilation (~4X in vessel diameters, p<0.01) and increased fluid accumulation (~3X in edematous regions, p<0.05) under lymphedema conditions. Lymphatic vascular dilation and edema were confirmed by fluorescent microscopy and histology.

Conclusions: OCT is an emerging non-invasive volumetric imaging modality that can be adapted to visualize lymphatic vasculature within normal and lymphedematous tissues without the use of vascular contrast agents. Furthermore, OCT has the ability to quantify the degree of vessel dilation and edema accumulation.

Citation Format: Jennifer Y. Kwan, Valentin Demidov, Blake Jones, Wei Shi, Justin Williams, Costel Flueraru, Kenneth W. Yip, Fei-Fei Liu, Alex Vitkin. Detection and quantification of lymphedema using optical coherence tomography lymphangiography [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 1644.
Abstract 1700: Excessive ROS in ovarian cancer cells impairs anti-tumor capacities of immune cells through exosomal miR-155/PD-L1 pathway
Authors: Li Xiang , Wang Gao-chan , He Jun
Published: September 2020

Background and Objective: Genetic and metabolic alterations in cancer cells promote the sustained oxidative stress as reflected by excessive production of reactive oxygen species (ROS). We and other groups have identified that the ROS-sensitive microRNAs regulate the cancer cell response to ROS. However, it has yet to be determined how these miRNAs convey the ROS signal to downstream effectors and thus produce biological consequences. Accumulative evidence indicates that tumor-derived exosomes are emerging mediators of tumorigenesis. In this study, we aimed to investigate how ROS convey the oncogenic signals through cellular interactions via exosomes within the tumor microenvironment.

Methods and Results: Through NanoString nCounter miRNA analysis, we identified miR-155-5p as the most abundant miRNAs enriched in the exosomes derived from epithelial ovarian cancer (EOC) cells treated with a ROS scavenger N-acetylcysteine (NAC). Mechanistically, tumor exosomal miR-155-5p was transferred to tumor-associated macrophages (TAM), directly targeting PD-L1 to inhibit the infiltration of TAM into tumor microenvironment. The role of tumor-derived exosomal miR-155-5p was determined by co-culture experiments and gain or loss of function studies in vitro, and confirmed in xenograft mouse models in nude mice and immunocompetent mice. In addition, we established a 3D-3-culture system to entail the co-culture of NAC-treated EOC cell spheroids, TAM and T cells. We found that CD8+ T-cell ratio was increased and production of IFN-γ, IL-2, and TNF-α was promoted. What's more, the immunocompetent EOC mouse model was established and treated with NAC-modified mouse ovarian cancer cell-released exosomes. The results showed that NAC-modified tumor growth was significantly reduced compared to control group. Further analysis showed that miR-155-5p increased in TAM, while PD-L1 expression levels decreased in macrophages from tumors and spleens. Moreover, the T cell function was also augmented as evaluated by the expression of Ki-67 and Granzyme B on CD8+ T cells.

Conclusion: In conclusion, our study reveals a novel mechanism underlying PD-L1 expression in EOC. Ovarian cancer cells with excessive ROS levels release exosomes that mediate the interaction between cancer cells and immune cells; it is this interplay that produces an immunosuppressive tumor microenvironment by upregulation of PD-L1 in macrophages, and eventually contributes to the progress of EOC through the ROS/miR-155-5p/PD-L1 pathway.

Citation Format: Xiang Li, Gao-chan Wang, Jun He. Excessive ROS in ovarian cancer cells impairs anti-tumor capacities of immune cells through exosomal miR-155/PD-L1 pathway [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 1700.
Abstract 1717: Targeted NanoSpheres (TNS): An effective, non-toxic nano-formulated therapeutic platform for broad use in cancer treatment
Authors: Kang Hyunggyoo , Nagy Jon , Triche Timothy
Published: September 2020

Current cancer therapy relies on the use of cytotoxic agents (‘chemotherapy') that was developed over 50 years ago. These agents are very effective, but efficacy is limited by off-target toxicity to normal tissues. It is widely recognized that chemotherapy could be far more effective if higher doses could be delivered to the tumor and not to normal tissues. By refining the early formulations and after extensive testing, we developed a novel targeted nanoparticle called Targeted NanoSpheres or TNS. We showed this novel nano-formulated platform can overcome many deficiencies of conventional nanoparticles and therefore has enhanced potential as an effective delivery vehicle for virtually any type of cancer for which a suitable tumor cell surface antigen is present. We are exploring the development of NV103, a first-in-class anti-CD99 TNS (CD99-TNS/Ir), for the treatment of Ewing Sarcoma (EWS), a highly lethal pediatric and young adult bone and soft tissue tumor driven by a fusion protein, typically EWS/FLI1 in over 85% of cases. This platform is also being investigated for other types of cancer that either express CD99, or other tumor antigens like GD2 and B7-H3. In addition, a variety of payloads, including small molecules chemotherapeutics like irinotecan and doxorubicin, as well as plasmids, siRNA, ASOs, and mRNA, have been successfully incorporated and effectively delivered with documented superior anti-tumor effect. NV103 itself showed significant anti-tumor effect in hepatocellular carcinoma and prostate tumor cell lines and xenografts. In all cases, systemic toxicity is minimized by use of targeting antibodies that enhance tumor delivery while minimizing systemic exposure of normal tissues.

Citation Format: Hyunggyoo Kang, Jon Nagy, Timothy Triche. Targeted NanoSpheres (TNS): An effective, non-toxic nano-formulated therapeutic platform for broad use in cancer treatment [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 1717.
Abstract 1720: Preclinical treatment of pancreatic ductal adenocarcinoma using targetable self-assembled gemcitabine nanoparticles
Authors: Fu Zhang
Published: September 2020

The prognosis of pancreatic ductal adenocarcinoma (PDAC) patients remains poor, and new effective therapies are urgently needed. Cytotoxic gemcitabine (GEM) is currently used as a standard-of-care drug for PDAC treatment but its efficacy is greatly compromised mainly due to quick metabolism in the body and drug resistance. Here we introduce a new simple approach to effectively deliver GEM to PDAC-derived tumors through the combination of a prodrug strategy, nanoparticle formulation, and PDAC-specific targeting strategy. By covalently conjugating GEM with a hydrophobic moiety (i.e. linoleic acid) via amide linkage, the resulting GEM prodrug was conferred with an amphiphilic feature and showed spontaneously self-assembling behavior in aqueous solution. The self-assembled nanoparticles were spherical and monodispersed in water with a diameter less than 100 nm. To optimize the nanoparticles for the in vivo use, DSPE-PEG2k was used to co-assemble with the prodrug. A PDAC-specific peptide was further decorated on the particle surface to endow the nanoparticles with PDAC targeting effect. In vitro assays on PDAC cell lines showed that the modified GEM nanoparticles exhibited superior efficacy than free GEM, and the conjugation of the targeting peptide significantly facilitated the internalization of the nanoparticles into the targeted cancer cells. Furthermore, in vivo evaluation in xenograft-bearing animal models derived from either established cancer cells or patient tumor tissues showed that these GEM nanoparticles were remarkably effective to inhibit tumor growth with alleviated side effects. These data confirmed the effectiveness of our self-assembling GEM prodrug strategy. Moreover, this cost-effective strategy can potentially be applied broadly to many other chemotherapeutic drugs that are insufficiently effective because of pharmacologic obstacles.

Citation Format: Zhang Fu. Preclinical treatment of pancreatic ductal adenocarcinoma using targetable self-assembled gemcitabine nanoparticles [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 1720.
Abstract 1808: Loss of BAP1/BRCA1 counteracts spindle assembly checkpoint activation by the anti-mesothelin antibody drug-conjugate anetumab ravtansine
Authors: Singh Anita , Luo Jin-li , Busacca Sara , Dawson Alan , Kolluri Krishna Kalyan , Janes Samuel , Fry Andrew , Sommer Anette , Fennell Dean
Published: September 2020

Mesothelioma is a rapidly lethal cancer with limited treatment options. Spindle poisons have demonstrated clinical usefulness in mesothelioma, however, predictive biomarkers are lacking. Anetumab ravtansine (ARav) is an antibody drug-conjugate (ADC) comprising a mesothelin targeting antibody coupled to the payload, DM4, a microtubule destabilizer. We have recently identified BRCA1 as a direct regulator of the spindle assembly checkpoint (SAC), which is activated by spindle poisons and mediates mitotic arrest. BAP1 is one of the most frequently inactivated tumor suppressors in mesothelioma and it interacts with and regulates expression of BRCA1. We therefore hypothesized that BAP1 inactivation may confer resistance to ARav, highlighting a potential opportunity to refine personalization with this ADC. Three primary cell lines were derived from mesotheliomas resected during pleurectomy decortication expressed mesothelin (MSLN) and further characterized via whole exome sequencing. Two of the three primary cell lines have BAP1 mutation (R417 frameshift (fs); E631fs) and show reduced BRCA1/MAD2 protein expression. MSLN expression was found to be independent of BAP1 inactivation status or BRCA1 expression level. BAP1 mutant primary cell lines exhibited resistance to ARav by overriding the mitotic arrest with continued cell cycle progression, indicating a defective SAC whereas the cell line with wt BAP1 underwent mitotic arrest followed by cell death. Increase in mitotic index was also observed in primary cell line with wt BAP1 following ARav treatment without any significant change in BAP1 mutant cell lines. Resistance to ARav in BAP1 deleted cell line NCI-H226 was reversed by stable expression of wildtype BAP1 in NCI-H226 indicating critical regulation of SAC by BAP1. Vinorelbine resistant cell line RVR with a defective SAC also exhibited resistance to ARav compared to the parental REN with no change in MSLN expression suggesting that an active SAC is required for prolonged mitotic arrest and subsequent cell death. Wildtype BAP1 and MSLN is critical for efficacy of ARav. Inactivation of BAP1 correlates with loss of BRCA1 expression and dysregulation of the SAC.

Citation Format: Anita Singh, Jin-li Luo, Sara Busacca, Alan Dawson, Krishna Kalyan Kolluri, Samuel Janes, Andrew Fry, Anette Sommer, Dean Fennell. Loss of BAP1/BRCA1 counteracts spindle assembly checkpoint activation by the anti-mesothelin antibody drug-conjugate anetumab ravtansine [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 1808.
Abstract 1821: Reversal of PD-L1-mediated chemoresistance by MCL-1/BCL-2 antagonism in colon cancer cells
Authors: Sun Lei , Patai Arpad , Qin Bo , Sinicrope Frank A.
Published: September 2020

Background: PD-L1 is an immune checkpoint protein that may display intrinsic, non-immune functions in human cancer cells. Previously, we discovered that PD-L1 is regulated by BRAFV600E via YAP and c-Jun, and whose deletion can confer chemoresistance due to a decrease in the BH3-only BIM protein in colorectal cancer (CRC) cells. Here, we hypothesize that release of pro-apoptotic BIM from its sequestration by MCL-1 or BCL-2 can overcomes drug resistance mediated by depletion of PD-L1.

Materials and Methods: Parental and PD-L1 knockout human (RKO) and murine (MC38) CRC cells with or without reconstituted PD-L1 were utilized. RKO cells with knockdown of MCL-1 or BCL-2 by shRNA were generated. Cells were treated with the MCL-1 selective inhibitor AZD5591 or the BCL-2/BCL-xL inhibitor ABT-263 alone or combined with the MEK inhibitor, cobimetinib. Apoptosis was measured by immunoblotting of cleaved caspase-3 and by Annexin V labeling using FACS. Clonogenic cell survival assay was performed. Protein/protein interaction was analyzed by immunoprecipitation assay.

Results: BIM induction by cobimetinib was insufficient to overcome chemoresistance due to loss of PD-L1. Using a potent and selective inhibitor of MCL-1 (AZD-5591), we demonstrate that concurrent antagonism of MCL-1 and MEK can overcome apoptosis resistance in both human and murine PD-L1 knockout CRC cells. Specifically, inhibition of MCL-1 by AZD-5591 or gene knockdown combined with cobimetinib suppresses clonogenic cell survival and enhances apoptosis in cells with PD-L1 knockout, wild type or those with reconstituted PD-L1. Mechanistically, the interaction of BIM with MCL-1 is increased in PD-L1 knockout (versus wild-type cells) in the presence of cobimetinib, and enhanced binding of BIM to MCL-1 is associated with BIM phosphorylation at Thr116. Targeting MCL-1 using AZD-5591 or MCL-1 shRNA releases BIM from its sequestration by MCL-1 and results in enhanced apoptotic signaling induced by cobimetinib to an equivalent extent in wild type and PD-L1 knockout cells with or without reconstitution of PD-L1. Similarly, targeting BCL-2 by ABT-263 or by BCL-2 shRNA releases BIM from its binding to BCL-2, and enhances apoptosis induction by cobimetinib independent of PD-L1 status. Enhanced binding of BIM to BCL-2 was also associated with BIM phosphorylation at Thr116.

Conclusion: Chemoresistance mediated by loss of PD-L1 can be reversed by the use of BH3 mimetic drugs combined with MEK inhibition in CRC cells to exploit their apoptotic dependency on BIM expression.

Citation Format: Lei Sun, Arpad Patai, Bo Qin, Frank A. Sinicrope. Reversal of PD-L1-mediated chemoresistance by MCL-1/BCL-2 antagonism in colon cancer cells [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 1821.
Abstract 1847: Phenotypic evaluation and discrimination of clinical JAK inhibitors for myeloproliferative disorders
Authors: Velichko Sharlene , Denker Sheryl P. , O'Mahony Alison
Published: September 2020

Small molecule inhibitors of Janus kinases (Jakinibs) represent a promising treatment for myeloproliferative disorders including polycythemia vera (PV), essential thrombocytosis (ET), and myelofibrosis (MF). Currently multiple JAK2 inhibitors (ruxolitinib, fedratinib, momelotinib, baricitinib) are either approved or in clinical development for the treatment of MF. The primary target of Jakinibs, the JAK-STAT pathway, plays a critical role in multiple cellular processes relevant for myelofibrosis, including survival, differentiation, and proliferation. However, all approved Jakinibs are reportedly promiscuous in that they inhibit kinases beyond the JAK family. Their overall clinical impact, therefore, reflects the sum of these target activities, as well as downstream secondary pharmacology relevant to both efficacy and safety. We evaluated these four Jakinibs using phenotypic profiling with the BioMAP® Diversity PLUS Panel to identify common versus differential activities. The BioMAP platform consists of human primary cell-based assay or systems designed to recapitulate key aspects of tissue and disease states. Distinct activity profiles were generated for each of the tested Jakinibs based on their impacts on 148 different biomarkers in 12 different model systems. Comparison of these profiles at concentrations relevant for clinical exposure levels revealed common activities including inhibition of T and B cell proliferation and decreased levels of inflammation and immune activation biomarkers. Together these activities represent a phenotypic signature related to shared primary JAK targets. In contrast, multiple differentiating activities were observed between the four drug profiles: baricitinib increased TNFα levels whereas momelotinib decreased TNFα in the LPS system modeling monocyte activation; fedratinib and momelotinib, but not ruxolitinib or baricitinib, inhibited endothelial cell proliferation; only momelotinib inhibited macrophage responses and decreased PGE2 levels. These data indicate that even within the specific JAK2 inhibitor drug class, individual entities have distinct modes of action relevant for clinical outcomes.

Citation Format: Sharlene Velichko, Sheryl P. Denker, Alison O'Mahony. Phenotypic evaluation and discrimination of clinical JAK inhibitors for myeloproliferative disorders [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 1847.
Abstract 1980: Multiplex digital PCR combined with melting curve analysis for detecting KRAS and BRAF mutations
Authors: Tanaka Junko , Nakagawa Tatsuo , Shiratori Akiko , Harada Kunio , Uematsu Chihiro , Takai Erina , Yachida Shinichi
Published: September 2020

Background

Digital PCR (dPCR) is a promising method for liquid biopsies and is able to quantify nucleic acids more sensitively than real-time PCR. However, dPCR has a large fluctuation in the fluorescence intensity of the droplets or wells due to insufficient PCR in the small partitions, limiting the multiplexing capability of using the fluorescence intensity. In this study, we propose a new measurement method that combines dPCR with melting curve analysis for highly multiplexed genotyping.

Methods

A sample is digitized into a silicon chip with up to ~104 wells in which asymmetric PCR was performed to obtain more single-stranded amplicons that are complimentary to molecular beacon probes. Fluorescence images were captured while controlling the temperature of the chip, and melting curve was measured for each well. Then, genotyping was performed by using the fluorescence intensity, dye color of the probe, and melting temperature (Tm). Because the Tm of the PCR products does not considerably depend on the amplification efficiency of PCR, genotyping accuracy is improved by using Tm values, enabling highly multiplexed genotyping.

Results

The concept was confirmed by measuring four common KRAS mutations (G12D, G12V, G12R, and G13D) and one common BRAF mutation (V600E). G12D, G12V and G12R-mutant KRAS alleles were detected with FAM-labeled probes, G13D-mutant KRAS allele and wildtype KRAS allele were detected with HEX-labeled probes, and V600E-mutant BRAF allele and wildtype BRAF allele were detected with CAL Fluor Red 610-labeled probes. The results showed that the peaks of the Tm distributions of the DNA groups were separated by more than 2°C. By combining the fluorescence intensities and the Tm values, the wildtype and mutant groups were clearly identified and able to be quantified.

Conclusion

We have proposed a new measurement method that combines dPCR with melting curve analysis, and successfully performed the multiplexed genotyping assay. To our knowledge, this is the first demonstration of genotyping of seven DNA groups with a single mutation of cancer-related genes by combining dPCR with melting curve analysis.

Citation Format: Junko Tanaka, Tatsuo Nakagawa, Akiko Shiratori, Kunio Harada, Chihiro Uematsu, Erina Takai, Shinichi Yachida. Multiplex digital PCR combined with melting curve analysis for detecting KRAS and BRAF mutations [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 1980.
Abstract 2000: Systemic and tumor associated IL-8 correlates with resistance to PD-L1 blockade
Authors: Yuen Kobe , Liu Lifen , Li Congfen , Rishipathak Deepali , Williams Patrick , Kadel Edward , Koeppen Hartmut , Madireddi Shravan , Keerthivasan Shilpa , Chen Ying-Jun , Modrusen Zora , Banchereau Romain , Leng Ning , Hegde Priti , Huseni Mahrukh , Mariathasan Sanjeev
Published: September 2020

Elevated plasma interleukin-8 (IL-8) is a poor prognostic factor for many cancers. However, the association of IL-8 with clinical outcomes to checkpoint inhibition has not been comprehensively evaluated in randomized studies. Moreover, the source of IL-8 and the underlying biology that influences resistance to immune checkpoint inhibitors remain unknown. Here we analyzed circulating IL-8 protein in plasma, and IL8 gene expression in peripheral blood mononuclear cells (PBMC) and tumors of patients treated with atezolizumab (anti-PD-L1 mAb), from multiple randomized trials in metastatic urothelial carcinoma (mUC) and metastatic renal cell carcinoma (mRCC). High levels of IL-8 in plasma, PBMCs and tumors, were associated with decreased efficacy in mUC and mRCC patients treated with atezolizumab, even in tumors that were classically CD8+ T cell inflamed. mUC patients treated with atezolizumab, but not with chemotherapy, who experienced an on-treatment decrease in plasma IL-8, exhibited improved overall survival. IL-8 is primarily expressed in circulating and intratumoral myeloid cells, and high IL8 expression was associated with the downregulation of the antigen presentation machinery in myeloid cells. A better understanding of IL-8-mediated myeloid inflammation in curtailing responses to checkpoint inhibitors is essential for developing new therapies for patients.

Citation Format: Kobe Yuen, Lifen Liu, Congfen Li, Deepali Rishipathak, Patrick Williams, Edward Kadel, Hartmut Koeppen, Shravan Madireddi, Shilpa Keerthivasan, Ying-Jun Chen, Zora Modrusen, Romain Banchereau, Ning Leng, Priti Hegde, Mahrukh Huseni, Sanjeev Mariathasan. Systemic and tumor associated IL-8 correlates with resistance to PD-L1 blockade [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 2000.
Abstract 2028: Recent trends in use of adjuvant chemotherapy in elderly stage II-III non-small cell lung cancer
Authors: Komiya Takefumi , Powell Emily , Guddati Achuta Kumar
Published: September 2020

Background: Although randomized trials demonstrated survival benefit of adjuvant chemotherapy, previous reports have suggested that its use in elderly populations for early stage non-small cell lung cancer (NSCLC) was infrequent. Oncologists often hesitate to offer adjuvant chemotherapy in resected early stage non-small cell lung cancer for elderly population. The status of adjuvant chemotherapy in this population is unknown for recent decades.

Materials and Methods: Using the Surveillance, Epidemiology, and End Results (SEER) database which represents about 28% of the US population, we examined the incidence of chemotherapy in resected stage II-III NSCLC between 2004 and 2015. Staging was determined according to the American Joint Committee on Cancer (AJCC) 6th version. Cases were grouped by age (20-69, 70-79, and 80+). Trends in use of chemotherapy by age group was assessed by univariate and multivariate analyses.

Results: A total of 35,009 were selected as surgically resected stage II-III NSCLC. Use of chemotherapy was 66.9%, 48.2%, 25.0% in age 20-69, 70-79, 80+, respectively. Multivariate analysis demonstrated that younger age (20-69) and recent year (2010-2015) of diagnosis were associated with increased use of chemotherapy. Chemotherapy use increased from 2004 to 2015 by 11.0%, 18.3%, and 11.3% in age 20-69, 70-79, 80+, respectively. In the age 70-79 group, increased use of chemotherapy was greater in stage II (24.3%) than stage III (14.1%). Five-year overall survival in age 70-79 group mildly increased by 7.6% from 2004 to 2011.

Conclusion: This study suggests that use of adjuvant chemotherapy in elderly population increased primarily in age 70-79. Few patients in age 80+ receive adjuvant chemotherapy even in recent years. This study not only informs oncologists of current status but also suggests further research on very elderly populations.

Citation Format: Takefumi Komiya, Emily Powell, Achuta Kumar Guddati. Recent trends in use of adjuvant chemotherapy in elderly stage II-III non-small cell lung cancer [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 2028.
Abstract 2038: Overall survival of glioblastoma patients treated by standard therapy in comparison to standard therapy plus tumor treating fields
Authors: Rulseh Aaron , Derner Adam , Sroubek Jan , Klener Jan , Vymazal Josef
Published: September 2020

Introduction: Glioblastoma multiforme (GBM) is the most common malignant primary intracranial tumor and traditionally has a median survival of only 10 to 14 months, with only 3 to 5% of patients surviving more than three years. Recurrence is nearly universal, and further decreases the median survival to only 5 to 7 months with optimal therapy. Tumor treating fields (TTFields; Optune, Novocure, Haifa, Israel) therapy is a novel treatment technique that has recently shown significant prolonged survival in GBM patients. This therapy is approved for the treatment of newly diagnosed and recurrent GBM and is based on the principle that low intensity, intermediate frequency alternating electric fields (100 to 300 kHz) have an anti-mitotic effect in specific cell types. The applied fields disrupt the mitotic spindle, microtubule assembly and the segregation of intracellular organelles during cell division, leading to apoptosis or mitotic arrest. We aimed to compare overall survival between patients treated with standard therapy and standard therapy plus TTFields at our institution.

Materials and Methods: A total of 34 patients (22 male, 15 female) diagnosed with GBM and treated by standard therapy plus TTFields (STDTh-TTF) at our institution were included. Standard therapy (STDTh) consisted of surgical resection, followed by combined radiotherapy and chemotherapy (Temozolomide); in 2 cases biopsy was performed in place of resection. The date of resection or biopsy was considered the entry date and was used in calculating survival, and ranged from June 2015 to July 2019. A matching control group treated by STDTh alone at our institution was assembled from our database, based primarily on date of resection or biopsy, and secondarily by age (1 subject underwent biopsy in place of resection). When assembling the control group, the investigators were blinded to survival outcome.

Results: Significantly greater overall survival was observed for the group treated by TTFields in addition to standard therapy (p = 0.005; Hazard ratio [HR] 0.28; 95% confidence interval [CI] 0.11-0.69). The groups were identical with respect to sex, and no differences with respect to age (p=0.12; STDTh-TTF mean 50.54 years, SD 10.3; STDTh mean 54.39 years, SD 9.3) or inclusion date (p=0.3) were detected.

Conclusions: Our initial results appear promising with respect to overall survival in patients undergoing TTFields treatment in addition to standard therapy.

Citation Format: Aaron Rulseh, Adam Derner, Jan Sroubek, Jan Klener, Josef Vymazal. Overall survival of glioblastoma patients treated by standard therapy in comparison to standard therapy plus tumor treating fields [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 2038.
Abstract 2044: Chemotherapy induced neutropenia in rats following administration of eflapegrastim or pegfilgrastim on the same day at three different timepoints and at 24 hours post-chemotherapy
Authors: Kim Yu-Yon , Kim Eunjung , Han Hyesun , Baek Seungjae , Chawla Shanta , Kolli Prasad , Bhat Gajanan , Lebel Francois
Published: September 2020

Background: Eflapegrastim is a long-acting G-CSF that is comprised of a G-CSF analog and a recombinant IgG4 Fc fragment conjugated at their N-termini via a short polyethylene glycol linker. Currently, G-CSF products are administered 24 hours after chemotherapy. We compared the duration of neutropenia (DN) after treatment with eflapegrastim and pegfilgrastim given on the same-day, at three different timepoints, and after 24 hours in chemotherapy-induced neutropenic rats.

Method: Neutropenia was induced in rats via intraperitoneal administration of cyclophosphamide and docetaxel (TC) chemotherapy. Rats (5 animals per group) were assigned to dose groups of either vehicle or equivalent doses (based on human clinical doses) of pegfilgrastim (103.3 µg/kg) or eflapegrastim (61.8 µg/kg as G-CSF) on the same day as chemotherapy at 0, 2, and 5 hours or 24 hours after chemotherapy. The primary endpoint was the difference in DN after TC treatment and administration of eflapegrastim or pegfilgrastim at the specified timepoints.

Results: In the vehicle group, the TC regimen induced neutropenia in all animals with an ANC nadir occurring on approximately Day 4 and a mean DN of nearly 7 days. The mean DN values for treatment with eflapegrastim on the same day, at 0, 2, and 5 hours, and after 24 hours were similar and the mean DN values for eflapegrastim were significantly lower than for pegfilgrastim at all dosing timepoints (Table).

Time after Chemotherapy(hours)Mean Duration of Neutropenia (days)Difference (95% CI)Eflapegrastim Time vs 24 hoursDifference (95% CI)EflapegrastimPegfilgrastim00.62.2-1.6 (-3.067, -0.133)0.4 (-0.08, 1.28)20.21.8-1.6 (-2.84, -0.36)0 (-0.19, 0.59)50.01.2-1.2 (-1.20, -1.20)-0.2 (0, 0)240.21.8-1.6 (-2.84, -0.36)NA

Conclusions: The DN for same-day treatment with eflapegrastim at 0, 2, and 5 hours was similar to that of the treatment after 24-hours. However, the mean DN values were significantly lower for eflapegrastim than pegfilgrastim at all time points. Further exploration of the potential advantage of same-day dosing with eflapegrastim in patients undergoing chemotherapy is warranted.

Citation Format: Yu-Yon Kim, Eunjung Kim, Hyesun Han, Seungjae Baek, Shanta Chawla, Prasad Kolli, Gajanan Bhat, Francois Lebel. Chemotherapy induced neutropenia in rats following administration of eflapegrastim or pegfilgrastim on the same day at three different timepoints and at 24 hours post-chemotherapy [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 2044.
Abstract 2057: Delivering on the Vision of Bench to Bedside: A Rare Disease Funding Community Collaboration to Develop Effective Therapies for Neurofibromatosis Type 1 Tumors
Authors: La Rosa Salvatore , Bakker Annette C. , Browder Vidya , Blakeley Jaishri O. , Verma Sharad K. , Wong Ling M. , Morris Jill , Bora Naba
Published: September 2020

The time from identifying a drug target to a new drug approval is often measured in decades and can take even longer for therapies to treat rare diseases. In fact, 95% of rare diseases do not have a specific therapy approved at all. Coordinated efforts to augment the drug development pipeline along with long-term and comprehensive support that enable scientific breakthroughs for rare diseases is possible, but it requires integration across multiple stakeholders. With the recently announced granted acceptance from the FDA to file status of the AstraZeneca and MSD (Merck) application for the MEK inhibitor selumetinib to treat plexiform neurofibromas in children, a major milestone towards the first-ever approved treatment for people with Neurofibromatosis Type 1 (NF1) associated tumors is closer than ever. This study analyzes the coordinated funding efforts of four federal and philanthropic organizations to advance drug development for NF1 associated tumors and discusses how these organizations collaborated and evolved practices to optimize funding and research support. This model could be replicated for other rare diseases if funders are able to take an active, coordinated approach to identify gaps, tools, and infrastructures most needed for that specific disease area.

Citation Format: Salvatore La Rosa, Annette C. Bakker, Vidya Browder, Jaishri O. Blakeley, Sharad K. Verma, Ling M. Wong, Jill Morris, Naba Bora. Delivering on the Vision of Bench to Bedside: A Rare Disease Funding Community Collaboration to Develop Effective Therapies for Neurofibromatosis Type 1 Tumors [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 2057.
Abstract 2059: Exploiting machine learning in local treatment strategies in cancer patients with bone metastases: A real world clinical trial
Authors: Zhao Hui , wang zhiyu , Sun jing , Gu yifeng , Yang mengdi , Yao guangyu
Published: September 2020

With rising life expectancy in cancer patients with bone metastases, the need for local treatment (LT) is expanding. Machine learning (ML) could create reasonable generalizations, the purpose of this article was to evaluate the use of ML model in LT strategies. Patients were treated by an interdisciplinary team in Shanghai Sixth People's Hospital. Visual analog scale (VAS) and Quality of Life (QoL) Questionnaire Bone Metastases Module scores were analyzed before, 1 week, 1, 3, and 6 months after treatments. ML models were used to calculate LT probability, and confusion matrix was used to calculate the accuracy, precision, recall and F1 score of models. ML models were further used to calculate pathological fracture (PF) probability in lung cancer patients. Of 386 patients enrolled between 2016 and 2017, 101 patients underwent LT. Significant improved VAS and pain domains scores were observed in 27 surgery patients at 1, 3 and 6m, while functional domains scores at 3 and 6m. All five scores improved significantly in 46 percutaneous osteoplasty patients at 1w, 1 and 3m. Significant improved VAS and pain domains scores were observed in 28 radiation patients at 1 and 3m, while functional domains scores at 3m. Compared with team decision-making, decision tree was superior to support vector machine and Bayesian neural networks in model building. The VAS scale, primary cancer, Frankel classification, Mirels score, C-terminal telopeptide of type I collagen (CTx), age, mid-fragment of osteocalcin (MID), character of bone metastases, CA153, and visceral metastases were included in the DT model. In 386 patients, the values of decision tree for the accuracy, precision, recall and F1 score were 86.53%, 78.44%, and 64.90% and 0.69 respectively. 124 lung cancer patients were used to calculate PF probability by decision tree. We also put driving gene mutation and five differentially expressed proteins into the model. The VAS scale, character of bone metastases, age, driving gene mutation, C-terminal telopeptide of type I collagen (CTx), mid-fragment of osteocalcin (MID) and enolase 1 (ENO1) were included in the DT model. The sensitivity, specificity and accuracy of DT model was 90.52%, 87.19% and 77.72%. Appropriate LT provided significant pain relief and improvement in QoL. The ML model is effective in helping physicians determine which patient may be the candidate for LT.

Citation Format: Hui Zhao, zhiyu wang, jing Sun, yifeng Gu, mengdi Yang, guangyu Yao. Exploiting machine learning in local treatment strategies in cancer patients with bone metastases: A real world clinical trial [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 2059.
Abstract 2094: Improving cancer cytogenetic case review with a new machine learning system
Authors: Roytman Megan , Wasnikar Viren , An Paul , Kashevan Raja , Verma Shalini , Shams Soheil
Published: September 2020

A novel machine learning system trained on The Cancer Genome Atlas (TCGA) data has been designed to help with classification, prognostic measures, and monitoring treatment responses in cancer management. The utility of this system will be demonstrated using a set of retrospective samples.

Cytogenetic analysis has long been utilized in clinical cancer care with conventional cytogenetic methods such as chromosomal banding and FISH still standard protocols for hematologic malignancies. With the advent of microarrays, smaller and more complex variants have been discovered in cancer samples but identifying the potential impact of many alterations such as copy number variation (CNV) and loss of heterozygosity (LOH) has not been fully exploited. A typical cancer cytogenetic report can list many tens of variants with unclear clinical implications.

To bridge this gap and assist with determining the clinical implications of such alterations, we have developed a novel machine learning system that has been trained using data from The Cancer Genome Atlas (TCGA). The TCGA data has been processed using the SNP-FASST2 segmentation algorithm followed by manual review for ploidy adjustment yielding high quality regions of CNV and LOH for over 6000 cases across 29 cancer types. Our novel neural network used this processed data to arrange samples based on their CNV and LOH profile across a two-dimensional map. The proximity of one sample to another on this map corresponds to its similarity in terms of CNV and LOH profile. To apply this system in a clinical setting, we have designed a case review system that aligns a new case with the learned cancer map to provide information on the similarity of the case with ones in TCGA. This enables rapid verification of tumor type or detection of a different origin for the sample than the one reported. In addition, the system can create a prognostic score using clinical survival data for the samples in TCGA.

Here we will demonstrate the utility of the system in classifying new cases against the learned cancer map and estimating prognostic measures.

Citation Format: Megan Roytman, Viren Wasnikar, Paul An, Raja Kashevan, Shalini Verma, Soheil Shams. Improving cancer cytogenetic case review with a new machine learning system [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 2094.
Abstract 2116: Strength of immune selection in tumors varies with sex and age
Authors: Castro Andrea , Pyke Rachel M. , Zanetti Maurizio , Carter Hannah
Published: September 2020

Individual MHC genotype constrains the mutational landscape during tumorigenesis. Immune checkpoint inhibition reactivates immunity against tumors that escaped immune surveillance in approximately 30% of cases. Recent studies demonstrated poorer response rates in female and younger melanoma patients. Although immune responses differ with sex and age, the role of MHC-based immune selection in this context is unknown. We found that tumors in younger and female individuals accumulated more poorly presented driver mutations than those in older and male patients, despite no differences in MHC genotype. Younger patients showed strongest effects of MHC-based driver mutation selection, with younger females showing compounded effects and nearly twice as much MHC-II based selection. This study presents the first evidence that strength of immune selection during tumor development varies with sex and age, and may influence the availability of mutant peptides capable of driving effective response to immune checkpoint inhibitor therapy.

Citation Format: Andrea Castro, Rachel M. Pyke, Maurizio Zanetti, Hannah Carter. Strength of immune selection in tumors varies with sex and age [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 2116.
Abstract 2148: Screening of cancer-reactive T cells in lymph nodes in colorectal cancer patients
Authors: Kiyotani Kazuma , Okamura Kazumi , Nagayama Satoshi , Nakamura Yusuke
Published: September 2020

Tumor draining lymph nodes (TDLNs) are located in the routes of lymphatic drainage from a primary tumor and have the highest metastasis in various types of solid tumors. TDLNs are considered as a tissue to activate the antitumor immunity, where antigen-specific effector T cells are generated and released to the blood stream. These effector T cells then infiltrate into the tumor site and attack cancer cells. However, it has not been investigated whether cancer-reactive T cells are present in lymph nodes. Through T cell receptor (TCR) repertoire analysis of cancer tissues and total 203 regional lymph nodes from 23 colorectal cancer patients, we previously found that regional lymph nodes, especially metastasis-positive lymph nodes, contain T cells with TCR shared with primary cancer tissues. In this study, we collected surgically-resected fresh cancer tissues and total 12 regional lymph nodes from 4 colorectal cancer patients. From these freshly-obtained cancer tissues and lymph nodes, we isolated single cells and then cultured T cells and patient-derived cancer cells. We co-cultured each of T cells from the 12 lymph nodes with autologous cancer cells, and examined their cancer-reactivity using IFN-γ ELISPOT assay. Six among the 7 metastasis-positive lymph nodes, shows significant production of INF-γ, suggesting the presence of cancer-reactive T cells in the lymph nodes. We also detected cancer-reactive T cells in 3 out of 5 metastasis-negative lymph nodes. In FACS analysis, we found the proportion of TIM3+ cells in CD8+PD1+ T cells were lower in T cells in lymph nodes compared to T cells in primary tumor. These data suggest that lymph nodes may be a good source of T cells for cancer immunotherapy.

Citation Format: Kazuma Kiyotani, Kazumi Okamura, Satoshi Nagayama, Yusuke Nakamura. Screening of cancer-reactive T cells in lymph nodes in colorectal cancer patients [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 2148.
Abstract 219: A high throughput single-cell workflow for paired genomic and phenotypic analysis
Authors: Dhingra Dalia , Mendez Pedro , Ooi Aik , Wang Shu , Gulati Saurabh , Sciambi Adam , Ruff Dave
Published: September 2020

Fusion gene detection has long been a focus of cancer research, when combined with mutations found in gDNA, can lead to a better understanding of disease progression. One example, BCR-ABL, a marker for CML and AML stem cells, is a target for tyrosine kinase inhibitor (TKI) treatments; however, there are mutations within BCR-ABL that evade TKIs and are selectively resistant to drug therapy. Single-cell technologies are now able to provide information into genomic DNA content, RNA expression, and protein surface markers, unmasked by the heterogeneity found in bulk data. However, multimodal analysis from the same single cell has not been straightforward to implement in a high throughput single-cell workflow. Here, we report the development of chemistries that enable analyses of both targeted genomic DNA and RNA sequencing from the same cell.

This workflow relies on the Tapestri platform, which uses a two-step microfluidic droplet system for the analysis of thousands of cells per run. The first droplet encapsulates each cell and releases the DNA while the second droplet introduces additional reagents for enzymatic manipulations on the cellular analytes. Novel primer designs are leveraged to capture both DNA and RNA and provide for independent barcoded sequence information. We first establish the utility of the Tapestri platform for accurate gene expression assessment with a targeted panel for breast cancer. For fusion gene transcript detection, panels were designed for AML and CML including primers targeting multiple potential variants of BCR-ABL transcripts. Cell lines were used to show high sensitivity and specificity of fusion sequence calls from RNA in cells where the expected SNVs, indels, and CNVs were also detected from gDNA. More complex targeted RNA and DNA panels, such as one for breast cancer with 35 gene expression amplicons and 88 genotyping amplicons, were also tested where we show agreement between cells clustered based on RNA expression and the cell assignment based on SNVs from gDNA.

This single-cell multimodal workflow on the Tapestri platform currently has the power to quantitatively link genotypic and phenotypic data from the same cell with future potential in biomarker identification and ultimately, to link to best-fit therapeutics.

Citation Format: Dalia Dhingra, Pedro Mendez, Aik Ooi, Shu Wang, Saurabh Gulati, Adam Sciambi, Dave Ruff. A high throughput single-cell workflow for paired genomic and phenotypic analysis [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 219.