bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 514: Correlation of sTNFR2 elevation and disease progression in cutaneous T cell lymphomas
Authors: Lee Amanda , Kim Youn , Khodadoust Michael , Faustman Denise L.
Published: June 2022

Tumor necrosis factor receptor 2 (TNFR2), a membrane receptor that is a part of the tumor necrosis factor (TNF) superfamily and one of two receptors that can bind to TNF-α, has been emerging in recent years as a promising cancer immunotherapy target. TNFR2 is commonly overexpressed in cancer cells and in the tumor microenvironment. It is also a key receptor in immune evasion, due to its abundance on the most immunosuppressive subset of Treg cells (CD4+Foxp3+), as well as its role in driving proliferation of regulatory T cells (Tregs) and tumor cells via activation of the nuclear factor kappa B (NF-κB) pathway. In environments with high cell proliferation and high surface densities, TNFR2 can be cleaved and shed as serum soluble TNFR2 (sTNFR2). In human cancers, high sTNFR2 has recently been associated with poor prognostic outcomes in colon cancer, esophageal cancer, endometrial cancer and non-Hodgkin’s lymphomas. Elevated sTNFR2 is also potentially associated with poor prognostic outcomes in cutaneous T cell lymphomas (CTCLs), a diverse group of lymphomas primarily composed of mycosis fungoides (MF) and Sézary syndrome (SS), with symptoms that primarily manifest in the skin. Given sTNFR2’s accessibility in serum and association with cancer prognosis, we investigated its role as a potential biomarker for MF and SS prognosis. We also evaluated whether the CTCL tumor type is mechanistically being driven by very suppressive Tregs and tumor cells with this oncogene. In our study, we measured baseline sTNFR2 levels from 31 serum samples across different cancer stages (Stage I-IV) of MF and SS in either treatment-naïve subjects or subjects free of any treatments for ≥3 months. Our initial data shows that sTNFR2 values correlate with levels of blood disease and increase as the cancer advances. These findings strengthen the possibility of using sTNFR2 as a biomarker in MF and SS and also support the possible mechanistic role of TNFR2 in disease pathogenesis. Analysis of the same serum samples for sTNFR1, a non-specific marker of inflammation and renal failure, confirm that the increases are exclusive to sTNFR2 and can indeed be attributed to MF and SS, rather than to non-specific inflammation. Further studies following these patients’ responses to their treatment plans (i.e., mogamulizumab, romidepsin) will help elucidate the potency of sTNFR2 in predicting response to treatment and cancer burden.

Citation Format: Amanda Lee, Youn Kim, Michael Khodadoust, Denise L. Faustman. Correlation of sTNFR2 elevation and disease progression in cutaneous T cell lymphomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 514.
Abstract 5166: Blood-based biomarker analysis for predicting efficacy of definitive concurrent chemoradiotherapy and durvalumab consolidation in patients with unresectable locally advanced non-small cell lung cancer
Authors: Park Cheol-Kyu , Park Hwa-Kyung , Oh Hyung-Joo , Cho Hyun-Ju , Oh In-Jae , Kim Young-Chul
Published: June 2022

Background: This study aimed to investigate the feasibility of using circulating tumor cells (CTCs) and peripheral blood cells (PBCs) as biomarkers for predicting the efficacy of concurrent chemoradiotherapy (CCRT) and durvalumab consolidation (DC) in patients with locally advanced non-small cell lung cancer (NSCLC).

Methods: We recruited patients diagnosed with unresectable stage III NSCLC who received definitive CCRT between March 2020 and March 2021. DC was permitted in patients who did not progress after CCRT and tumor PD-L1 ≥1%. Blood samples were collected before (C0) and after CCRT (C1) to calculate PBC counts and analyze CTCs. CTCs, isolated using CD-PRIME࣪ system, exhibited EpCAM/CK+/CD45− phenotype in BioViewCCBS࣪.

Results: A total of 50 patients were enrolled and 23 patients received DC. The median duration of follow-up was 16.3 months, and the median progression-free survival (PFS) was not significantly different between patients with and without DC (13.1 vs. 13.5 months; p=0.355). In overall, patients with higher platelets (PLThi, >252x103/uL) at C1 had worse median PFS than those with lower platelets (PLTlo, ≤252x103/uL) (5.9 vs. 15.1 months; p<0.001). In DC group, patients with residual CTC clusters after CCRT (C1) had worse median PFS than those without detectable CTC cluster (9.5 vs. not reached; p=0.034). In multivariate analysis, PLThi at C1 was an independent factor for PFS (hazard ratio [HR] 4.12, 95% confidence interval [CI] 1.77-9.61; p=0.001), and patients with DC who had PLThi and residual CTC clusters at C1 showed the worst PFS (2.6 months, HR 44.40; p=0.001), even worse than those without DC who had PLThi (5.9 months, HR 15.65; p=0.002).

Conclusions: Comprehensive analysis of CTC and PBC counts before and after CCRT, especially CTC clusters and platelets at C1, demonstrated they might be biomarkers for predicting survival. This finding could aid in risk stratification of patients with unresectable stage III NSCLC who are eligible for DC after definitive CCRT.

Citation Format: Cheol-Kyu Park, Hwa-Kyung Park, Hyung-Joo Oh, Hyun-Ju Cho, In-Jae Oh, Young-Chul Kim. Blood-based biomarker analysis for predicting efficacy of definitive concurrent chemoradiotherapy and durvalumab consolidation in patients with unresectable locally advanced non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5166.
Abstract 5167: Enhanced isolation of circulating tumor cells from leukapheresis material enabled by Akadeum microbubbles
Authors: Grimley Edward , Roussey Jon , Ku Chia , Livi Carolina , Petlakh-Co Nadia , Wegner Casey , McNaughton Brandon H.
Published: June 2022

Circulating tumor cells (CTCs) have shown incredible promise in clinical and diagnostic applications. CTC-based liquid biopsies can aid in earlier diagnosis, identify the stage of disease, guide therapeutic aggressiveness, and be used to monitor response to treatment. While CTCs hold incredible promise in these areas, there are inherent limitations that hinder realization of the full potential of CTC-based liquid biopsy tests. The rarity of CTCs is one of the most significant factors limiting clinical utility. A standard 10-mL blood draw may simply not contain the number of CTCs needed to yield meaningful results. One method to analyze greater blood volume is by leukapheresis. While clinical leukapheresis can enrich mononuclear cells, including CTCs, from almost the entire volume of a patient’s blood, most downstream CTC enrichment methods are not compatible with leukapheresis material as a starting sample. To overcome these compatibility issues, Akadeum Life Sciences has developed a novel and proprietary Buoyancy Activated Cell Sorting (BACS™) microbubble platform that can be functionalized to selectively enrich biological samples for target cell populations using a fast, easy protocol that is exceptionally gentle on delicate cells of interest. Akadeum’s low-density BACS microbubbles are mixed into the sample where they quickly engage their targets and naturally float them to the surface for removal. This straightforward approach is elegant in simplicity and powerful in application, offering some of the fastest and most gentle isolations available. In this study, we demonstrate a protocol for the depletion of white blood cells (WBCs) & residual red blood cells (RBCs) from leukapheresis material via microbubbles. This novel protocol reveals a > 99% combined depletion of residual RBCs and WBCs, providing over 120-fold enrichment of tumor cells. Importantly, this protocol delivers exceptional yield with > 99% retention of the target tumor cells. This optimized workflow delivers an enriched and viable population of untouched and unaltered CTCs for downstream use.

Citation Format: Edward Grimley, Jon Roussey, Chia Ku, Carolina Livi, Nadia Petlakh-Co, Casey Wegner, Brandon H. McNaughton. Enhanced isolation of circulating tumor cells from leukapheresis material enabled by Akadeum microbubbles [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5167.
Abstract 5169: Sentinel lymph node gene expression signature predicts survival outcome in cutaneous melanoma
Authors: Karapetyan Lilit , Gooding William , Yang Xi , Knight Andrew , Li Aofei , Sander Cindy , Panelli Monica , Storkus Walter J. , Tarhini Ahmad A. , Kirkwood John M.
Published: June 2022

Background: Sentinel lymph node status is an important prognostic factor for melanoma progression. We sought to develop a sentinel lymph node gene expression signature score predictive of disease progression in patients with cutaneous melanoma.

Methods: Cases were retrieved from University of Pittsburgh protocol “Sentinel Node and Non-Sentinel Lymph Nodes (SLN and non-SLN) Procurement from Melanoma Subjects for Molecular Profiling.” Gene expression profiling was performed on SLN biopsies using U133A 2.0 Affymetrix gene chips. The top 25 genes associated with progression-free survival (PFS) were selected and a penalized regression function was used to select genes with a non-zero coefficient. A risk score was calculated from a 12 gene signature. A proportional hazards regression model was used to evaluate the association between clinical covariates, gene signature score, and PFS.

Results: In total, 45 patients were included [female=18 (40%), median age (range)= 56 (16-81) years]. The median Breslow thickness (IQR) was 4.1 (3.3-6.0) mm, ulceration was present in 29 (67%) cases, and 25 (56%) lesions had nodular histology. Among patients, 23 (51%) had a positive SLN via H&E. Twenty-one (46.7%) patients developed disease recurrence/progression with a median follow-up of 8.3 years. The median time to progression was 9.6 years. Among the top 25 genes, 12 non-zero penalized coefficients were estimated (CLGN, C1QTNF3, ADORA3, ARHGAP8, ZNF223, PDE6G, CXCL3, CHRFAM7A, HLA-DRB-decreased risk, DCTN1, ASPSCR1, HEXIM1-increased risk). A 12-gene signature score was significantly associated with PFS (p<0.0001) and produced a bootstrap cross-validated C index of 0.888. Stratifying by SLN status the gene signature risk score remained highly significant (p<0.0001). In univariate analysis, Breslow thickness (HR=1.41), presence of ulceration (HR=3.61), SLN positivity (HR=2.50), and acral-lentiginous histology compared to nodular (HR=4.07) were each significantly associated with PFS (all p<0.05). After simultaneously adjusting for all of these factors plus the gene signature, the 12-gene score remained a significant predictor for PFS (p<0.0001).

Conclusion: A SLN 12-gene signature risk score is associated with melanoma progression regardless of SLN status and can be used as a prognostic factor for PFS of patients. External validation of the gene signature is warranted.

Citation Format: Lilit Karapetyan, William Gooding, Xi Yang, Andrew Knight, Aofei Li, Cindy Sander, Monica Panelli, Walter J. Storkus, Ahmad A. Tarhini, John M. Kirkwood. Sentinel lymph node gene expression signature predicts survival outcome in cutaneous melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5169.
Abstract 517: Protein disulfide isomerase as a predictive biomarker for non-muscle invasive bladder cancer recurrence
Authors: Yuen Kit L. , Wu Chia-Hao , Silvers Christopher R. , Steinmetz Alexis R. , Miyamoto Hiroshi , Messing Edward M. , Lee Yi-Fen
Published: June 2022

INTRODUCTION AND OBJECTIVE: High recurrence rates found in non-muscle invasive bladder cancer (NMIBC) pose a financial and psychological burden for patients. We demonstrated that bladder cancer (BC) cells release protein disulfide isomerase (PDI), a redox-regulated ER-resident protein, via extracellular vesicles (EVs) when experiencing high levels of ER stress for their own survival. Chronic exposure to PDI-enriched BC EVs induced malignant transformation in recipient non-transformed cells. We hypothesized that the dynamic redox state in BC cells determines PDI cellular localization and tumors with elevated PDI preferentially incorporate PDI into EVs for removal. Furthermore, PDI expression levels and subcellular localization in tumors from NMIBC patients may predict NMIBC recurrence.

METHODS: Retrospective chart review identified 122 patients with newly diagnosed low-grade NMIBC classified into 2 groups: recurrent (N=55) and non-recurrent (N=67) during follow-up period. Formalin fixed paraffin embedded tumor specimens from index transurethral resection of bladder tumor (TURBT) were obtained and a tissue microarray (TMA) was constructed. Immunofluorescent staining of TMA sections was conducted using antibodies against PDI and BiP, an ER stress marker. Stained tissues were photographed and tumor regions in each field were manually masked and confirmed by a genitourinary pathologist. Within each tumor region, total antibody labeling was determined by measuring the mean pixel intensity values with NIH ImageJ/Fiji. Manders coefficient was used to assess the colocalization of epitopes using JACoP plugin in ImageJ.

RESULTS: Tumor tissues from patients that recurred expressed significantly higher levels of PDI and BiP. Moreover, high PDI and BiP expression predicted worse recurrence-free survival. These data suggest that tissues under higher levels of ER stress—as indicated by higher levels of PDI and BiP expression—have a higher risk of recurrence. Significantly higher levels of PDI were detected in non-ER cytosol in recurrent cohort tumors.

CONCLUSIONS: This study provides a novel mechanism for BC recurrence and valuable clinical predictive biomarkers for NMIBC recurrence and progression. Future clinical applications targeting this pathway may be an avenue to prevent NMIBC recurrence.

Citation Format: Kit L. Yuen, Chia-Hao Wu, Christopher R. Silvers, Alexis R. Steinmetz, Hiroshi Miyamoto, Edward M. Messing, Yi-Fen Lee. Protein disulfide isomerase as a predictive biomarker for non-muscle invasive bladder cancer recurrence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 517.
Abstract 5198: Dynamics of circulating immune cells during chemoradiotherapy in patients with locally advanced non-small cell lung cancer support earlier administration of anti-PD-1/PD-L1 therapy
Authors: Kim Kyung Hwan , Pyo Hongryull , Oh Dongryul , Noh Jae Myoung , Ahn Yong Chan , Yoon Hong In , Lee Jiyun , Park Sehhoon , Jung Hyun-Ae , Sun Jong-Mu , Lee Se-Hoon , Ahn Jin Seok , Park Keunchil , Ku Bo Mi , Ahn Myung-Ju , Shin Eui-Cheol
Published: June 2022

Background: Chemoradiotherapy (CRT) followed by consolidation immune checkpoint inhibitors (ICIs) significantly improves survival in unresectable locally advanced non-small cell lung cancer (LA-NSCLC). However, the optimal sequence for CRT and ICIs has not yet been established. We investigated the dynamics of peripheral blood immune cells during CRT to determine the best sequence for treatment.

Methods: Peripheral blood samples were prospectively collected pre-treatment, weekly during CRT for 6 weeks, and 1 month post-treatment in 24 patients with LA-NSCLC who received definitive CRT. Immune cell analysis was performed by flow cytometry. Ex vivo PD-1 blockade assays were performed by IFN-γ intracellular cytokine staining.

Results: Lymphopenia was prominently observed during CRT and mostly recovered 1 month post-CRT. Robust proliferation of CD8+ T cells was induced, peaking in the last week during CRT and decreasing post-CRT. The robust proliferation of CD8+ T cells led to an increase in the frequency of CD28-CD57+ replicative senescent and terminally differentiated cells post-CRT. Tumor-reactive CD8+ T cells increased during CRT and peaked in the last week. One month post-CRT, the frequency of tumor-reactive CD8+ T cells decreased and TOXhiTCF1lo terminally exhausted CD8+ T cells significantly increased. Anti-PD-1-induced functional restoration of PD-1+CD8+ T cells was maximized in the last week of CRT and significantly decreased post-CRT.

Conclusions: The findings suggest that earlier administration of PD-1 blockade may be associated with superior efficacy compared to delayed administration after completion of CRT. These findings provide an immunological rationale for optimal timing of combining ICIs with CRT in clinical trials.

Citation Format: Kyung Hwan Kim, Hongryull Pyo, Dongryul Oh, Jae Myoung Noh, Yong Chan Ahn, Hong In Yoon, Jiyun Lee, Sehhoon Park, Hyun-Ae Jung, Jong-Mu Sun, Se-Hoon Lee, Jin Seok Ahn, Keunchil Park, Bo Mi Ku, Myung-Ju Ahn, Eui-Cheol Shin. Dynamics of circulating immune cells during chemoradiotherapy in patients with locally advanced non-small cell lung cancer support earlier administration of anti-PD-1/PD-L1 therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5198.
Abstract 5212: Braf-altered microsatellite-stable metastatic colorectal cancer: A niche for immunotherapy
Authors: Kuan Feng-Che , Shi Chung-Sheng , Wu Yu-Ying , Lin Meng-Hung
Published: June 2022

Background: The successful inhibition of the PD-1/PD-L1 axis and CTLA-4 has opened the era of immuno-oncology, however only a few patients with metastatic colorectal cancer (mCRC) and microsatellite instability (MSI, ~5%) derive survival benefits from these treatments. BRAF genomic alterations, mainly V600 point mutations, comprise 10-15% of cases and have a poorer prognostic phenotype.

Method: The public domain genomic database (via cBioPortal and AACR Project GENIE) was used as an external control to explore the genomic make-up of BRAF-mutated and BRAF-wild mCRC tumors. The Wilcoxon sum rank test was used for the genome-wide association analysis, and the Kaplan-Meier method was used to estimate survival. Besides, clinical observations on extraordinary responses to regorafenib plus nivolumab therapy in two patients with microsatellite stable (MSS) mCRC with BRAF genomic alterations were reported.

Result: The CRC cohort comprised 2,936 patients and the immune checkpoint inhibitor (ICI) treated cohort comprised 1,661 patients, of which 110 (6.6%) were CRC patients. BRAF genomic alterations (CRC versus ICI cohort: 12.4% versus 11.8%, respectively) were significantly associated with POLE and PRKDC gene mutations (p<0.001). The genomic signatures, such as higher mutation count (MC) (log2 [6.54 versus 3], Wilcoxon p<10-10) and lower fraction genome altered (FGA) (log2 [-3.62 versus -2.22], Wilcoxon p=2.08e-9) were similar to that of POLE and PRKDC in the CRC cohort. Also, BRAF genomic alterations are associated with higher mutation counts in not only MSI but MSS cohorts (p=0.033 and p<0.001, respectively). In the ICI cohort, BRAF-altered colorectal tumors were associated with a higher TMB (43.3 versus 7.81, Wilcoxon p=0.016). In the CRC cohort, BRAF-altered tumors were associated with poorer overall survival (median 38.04 versus >80 month, log rank p<0.0001); and in the ICI cohort, BRAF-altered colorectal tumors were associated with a trend of better overall survival after ICI treatment (median >80 versus 15 months, log rank p=0.121)

Conclusion: BRAF-mutated malignancies represent a sub-population of MSS CRC with higher TMB, which could benefit from a combination of regorafenib and nivolumab therapy.

Citation Format: Feng-Che Kuan, Chung-Sheng Shi, Yu-Ying Wu, Meng-Hung Lin. Braf-altered microsatellite-stable metastatic colorectal cancer: A niche for immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5212.
Abstract 5226: The PI3K inhibitor Paxalisib combines synergistically with the pan-Raf inhibitor TAK580 (DAY 101) to extend survival in orthotopic xenograft models of atypical teratoid/rhabdoid tumors
Authors: Malebranche Kristen , Findlay Tyler , Eberhart Charles , Raabe Eric , Rubens Jeffrey
Published: June 2022

Atypical teratoid/rhabdoid tumors (AT/RT) are aggressive pediatric brain tumors and the most common malignant brain tumors of infancy. The four-year event-free survival rate is only 37%. For patients with relapsed AT/RT, there are limited treatment options, but novel precision therapies may help improve survival for patients with this deadly disease. We have previously identified strong activation of the PI3K-AKT-mTOR (mTOR) signaling pathways in AT/RT. Paxalisib is a highly brain-penetrant PI3K inhibitor acting upstream of mTOR. We find that Paxalisib slows tumor growth in orthotopic xenograft models of AT/RT and extends median survival from 40 to 54 days (p=0.001, log-rank test). RNASeq after mTOR pathway inhibition identifies reflexive activation of the MAPK pathway as a possible mechanism of therapy resistance (KEGG pathway analysis). TAK580 (DAY 101) is a highly brain penetrant, pan-RAF kinase inhibitor, currently in clinical trials in pediatric low grade gliomas (NCT03429803, NCT04775485). Paxalisib combines synergistically with TAK580 to reduce AT/RT growth and viability (Bliss synergy score 24.22). A pilot study suggests combination therapy is well tolerated in mice bearing AT/RT orthotopic xenografts and slows tumor growth compared to Paxalisib alone and vehicle controls. This novel combination of precision therapies can translate into a clinical trial aimed at improving survival in this deadly disease.

Citation Format: Kristen Malebranche, Tyler Findlay, Charles Eberhart, Eric Raabe, Jeffrey Rubens. The PI3K inhibitor Paxalisib combines synergistically with the pan-Raf inhibitor TAK580 (DAY 101) to extend survival in orthotopic xenograft models of atypical teratoid/rhabdoid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5226.
Abstract 5227: Combination therapy activating the integrated stress response synergistically suppresses proliferation and induces apoptosis in diffuse intrinsic pontine glioma
Authors: Novak Orlandi V. , Arnold Antje , Eberhart Charles , Raabe Eric H.
Published: June 2022

Diffuse intrinsic pontine glioma (DIPG) is a lethal pediatric brain tumor. Radiation, the standard of care, extends life for 6-18 months but has never cured a patient. Therefore, the discovery of novel treatments is imperative. DIPG tumors have elevated baseline activation of the integrated stress response (ISR), an evolutionarily conserved system that allows cells to tolerate various forms of stress. Increased expression of activating transcription factor 4 (ATF4) indicates activation of the ISR. Low levels of ATF4 protect cells from stress, while sustained high-levels of ATF4 result in cell death. Because DIPG has a high baseline level of ATF4, we hypothesized that the ISR activators Sal003 and ONC201 would synergize and kill DIPG cells. After determining the IC25 of Sal003 and ONC201, we treated three patient-derived cell lines: JHH-DIPG1, SF-7761, and JHH-DIPG16A with low micromolar doses. To measure proliferation, we performed immunofluorescence staining for bromodeoxyuridine (BrdU) incorporation. The combination treatment significantly reduced BrdU incorporation (JHH-DIPG1 p=0.0026, SF-7761 p=0.0002, JHH-DIPG16A p<0.0001 by ANOVA and Dunnett’s multiple comparisons test compared to DMSO control). In all three cell lines, the combination also significantly reduced proliferation compared to monotherapy. We next measured apoptosis by staining for cleaved caspase-3 (CC3) and performing western blots for cleaved PARP. The combination of Sal003 and ONC201 significantly increased apoptosis as measured by CC3 immunofluorescence in comparison to DMSO (JHH-DIPG1 p<0.0001, SF-7761 p<0.0001, JHH-DIPG16A p<0.0001 by ANOVA and Dunnett’s multiple comparisons test compared to DMSO control). In all cell lines, combination therapy significantly increased CC3 positivity compared to single treatment. Western blots for cleaved PARP expression detected induction of apoptosis in all three cell lines treated with the combination over DMSO and monotherapy treated cells. In JHH-DIPG1 and SF-7761, the combination increased ATF4 expression. Since Sal003 is not yet available for clinical testing in humans, we will next investigate treatment with ONC201 and the well-tolerated ATF4 inducer fenretinide. The combination of ONC201 with another ISR activating agent has the potential to serve as a therapy for DIPG.

Citation Format: Orlandi V. Novak, Antje Arnold, Charles Eberhart, Eric H. Raabe. Combination therapy activating the integrated stress response synergistically suppresses proliferation and induces apoptosis in diffuse intrinsic pontine glioma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5227.
Abstract 5249: Efficacy and safety of apatinib in treatment of unresectablehepatocellular carcinoma: A real-world study
Authors: Liu Jun , Li Guangbing , He Qiang , Sun Shijun , Zhao Chuanlin , Sun Chuandong , Zhang Bo , Cui Yi , Ye Yongqiang , Zhi Xuting , Li Shiping , Li Shiping , Pan Guozheng , Ma Changlin , Hu Jinhua , Li Jun , Wang Suling , Qiao Yuangang , Ren Wanhua , Meng Qi
Published: June 2022

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in China. Apatinib is a small molecule TKI inhibitor. Its efficacy and safety for unresectable HCC patients have been demonstrated in randomized controlled clinical studies. However, the efficacy and safety of apatinib in real world is lacking. We are aiming to evaluate the efficacy and safety of apatinib in real world.MethodsPatients of older than 18 years with unresectable HCC confirmed by IHC (Immunohistochemistry) or CT and MR Imaging were enrolled in the study. All patients received apatinib monotherapy or combined with other treatments including TACE and chemotherapy. The enrolled patients were treated until disease progression, death, or severe intolerable toxicity, etc. The primary endpoint was overall response rate (ORR), and secondary endpoints were disease control rate (DCR), progression-free survival (PFS), and overall survival (OS). The adverse events (AEs) were also recorded.ResultsFrom February, 2019 to May, 2020, a total of 233 patients were enrolled. Among them, 66 (28.33%) patients received apatinib monotherapy and 167 (71.67%) patients received combination therapy. Patients with a history of hepatitis accounted for 70%. 8 patients achieved complete response (CR), 64 patients achieved partial response (PR), 120 patients achieved stable disease (SD), and 41 patients had progressive disease (PD), illustrating an ORR of 30.90% and a DCR of 82.40%. Median PFS and OS were 6.93 months (95% CI, 6.19-7.68 months) and 11.36 months (95% CI, 9.96-12.77 months), respectively. The safety profile indicated that the most common drug-related adverse events were hypertension (37.90%), hand-foot-skin reaction (27.39%), fatigue (19.11%), thrombocytopenia (16.02%). Grade 3/4 treatment related AEs included thrombocytopenia (4.30%), hypertension (3.82%), neutropenia (2.73%). After symptomatic treatment, all the adverse events were treated properly, and no unexpected adverse events were observed. Conclusions In current study, apatininb showed good tolerance with acceptable toxicity and high tumor response rate that translated into promising PFS and OS in unresectable HCC patients. ChiCTR1900021822

Citation Format: Jun Liu, Guangbing Li, Qiang He, Shijun Sun, Chuanlin Zhao, Chuandong Sun, Bo Zhang, Yi Cui, Yongqiang Ye, Xuting Zhi, Shiping Li, Shiping Li, Guozheng Pan, Changlin Ma, Jinhua Hu, Jun Li, Suling Wang, Yuangang Qiao, Wanhua Ren, Qi Meng. Efficacy and safety of apatinib in treatment of unresectablehepatocellular carcinoma: A real-world study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5249.
Abstract 5254: Association of USP6 mutation with tumor mutation burden and survival in melanoma patients treated with immune checkpoint inhibitors
Authors: Long De , Fan Dandan , Chen Yaoxu , Huang Mengli
Published: June 2022

Background: Melanoma is a serious skin cancer. Immune checkpoint inhibitors (ICIs) including pembrolizumab, nivolumab, ipilimumab have been approved by FDA. However, ICIs demonstrate antitumor effects only in a fraction of patients and research exploring the association between gene mutation and clinical benefit is limited. USP6 (Ubiquitin-specific protease 6) mutation rate is high in melanoma. Studies have shown that USP6 functions as a tumor suppressor in Ewing Sarcoma through the activation of NK cells and macrophages, which indicates that USP6 activity may create a "hot" tumor microenvironment in immunotherapy. The association between USP6 mutation and survival in melanoma is unknown.

Methods: The association between USP6 mutation with TMB and survival data was analyzed in melanoma patients from the public immunotherapy-treated cohort called Melanoma.Allen2015.WES.110, which worked as training cohort while another cohort called Melanoma.Hugo2016.WES.38 worked as validation cohort. Wilcoxon test was used for the comparison of TMB. OS (Overall survival) analysis were conducted in the public cohort using Kaplan-Meier curves and log-rank tests. In addition, the genomic and immune cell infiltration data of 466 patients with Skin Cutaneous Melanoma was obtained from TCGA and the correlation analysis was further analyzed by CIBERSORT. Statistical significance was set at p=0.05.

Results: In the training cohort, 10.9% (12/110) melanoma patients harbored USP6 mutation. USP6 mutation is associated with higher TMB (p=0.0007) and results in significantly longer OS (21.3 vs 8.1 months; HR, 0.44; p=0.049).Meanwhile, the validation cohort shows that USP6 mutation results in higher TMB without significant difference (p=0.08) and significantly longer OS (31.5 vs 14.4 months; HR, 0.14; p=0.0026).Furthermore, the correlation analysis between immune infiltration and USP6 mutation status in melanoma shows that both M1 macrophages and follicular helper T cells increased significantly (p=0.02, p=0.033), while differential neutrophil decreased significantly (p=0.048).

Conclusions: This study shows that USP6 mutation may serve as a potential positive biomarker of ICIs in melanoma since it correlated with higher TMB, the up regulation of M1 macrophages, follicular helper T cells and the down regulation of differential neutrophil.

Citation Format: De Long, Dandan Fan, Yaoxu Chen, Mengli Huang. Association of USP6 mutation with tumor mutation burden and survival in melanoma patients treated with immune checkpoint inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5254.
Abstract 5266: Prognostic role of molecular subclassification based on mutational profiling and tumor-associated neutrophil status in stage III colon cancer
Authors: Chan CY , Lo AWI , Lam KO , Ng HY , Chen CC , Lung ML , Ko JMY
Published: June 2022

The traditional prognostic prediction of colon cancer (CC) is based on the tumor-node-metastasis (TNM) system. This, however, is not adequate for clinical decision making due to the existence of inter-patient variations and heterogeneity of underlying molecular pathways. We aim to investigate the prognostic value of molecular subclassification utilizing i) tumor-associated neutrophil (TAN) status by immunohistochemical (IHC) staining; ii) microsatellite instability (MSI) and elevated microsatellite alteration at selected tetranucleotide repeats (EMAST) status by PCR-based microsatellite typing; and iii) next-generation sequencing (NGS) mutational profiling by deep sequencing of a 77-gene oncology panel. In the discovery phase, 130 pairs of archived normal and tumor formalin-fixed paraffin-embedded (FFPE) specimens of stage III CC patients were retrospectively recruited from Queen Mary Hospital. CC patients will be categorized into different molecular subclasses (MSCs) based on the status of TAN/MSI/EMAST/NGS mutational profiling molecular biomarkers. Kaplan-Meier analysis, log rank test, univariate and multivariate Cox proportional hazards regression analysis will be used for estimation of progression-free survival (PFS) and overall survival (OS) between the two groups of CC patients. We expect our data to provide useful information for improved stratification of stage III CC patients into high or low risk of recurrence and survival.

Acknowledgement: The Health and Medical Research Fund (07181536) to JMYK.

Citation Format: CY Chan, AWI Lo, KO Lam, HY Ng, CC Chen, ML Lung, JMY Ko. Prognostic role of molecular subclassification based on mutational profiling and tumor-associated neutrophil status in stage III colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5266.
Abstract 5311: Auranofin sensitizes paclitaxel induced apoptotic cell death through oxidative mechanism in breast cancer cells
Authors: Prasad N. Rajendra , Natarajan Deepika
Published: June 2022

Breast cancer cells exhibit several basal redox systems, thus poorly responding to oxidative cell death. These intrinsic redox statuses capitalize on the efficacy of chemotherapeutic drugs. Auranofin, an FDA-approved gold compound documented with effective pharmacokinetics and safety profiles in humans, has recently been repurposed for anticancer activity. This study examined the paclitaxel sensitizing effect of auranofin by targeting redox balance in the MDA-MB-231 and MCF-7 breast cancer cell lines. We observed that auranofin inhibits thioredoxin reductase activity (TrxR) in these breast cancer cells and collapses the cellular antioxidants status, including the reduced activities of superoxide dismutase, catalase, and glutathione peroxidase. Furthermore, it has been noticed that auranofin augmented paclitaxel mediated cytotoxicity in a concentration dependant manner in both the MDA-MB-231 and MCF-7 cell lines. Furthermore, auranofin increased intracellular ROS levels (DCFH-DA staining) and altered mitochondrial membrane potential (Rhodamine-123 staining) in these breast cancer cell lines. The proapoptotic markers such as p53, Bax, and caspase-3 and caspase-9 expressions were higher in auranofin plus paclitaxel treated breast cancer cells when compared to paclitaxel alone treatment. Thus, the present results illustrate the chemosensitizing property of auranofin in MDA-MB-231 and MCF-7 breast cancer cell lines.

Citation Format: N. Rajendra Prasad, Deepika Natarajan. Auranofin sensitizes paclitaxel induced apoptotic cell death through oxidative mechanism in breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5311.
Abstract 5314: Selinexor, a selective inhibitor of nuclear export (SINE) compound, enhances the anti-tumor activity of Eribulin in leiomyosarcoma
Authors: Mittal Sonam , Pulikkal Ishaque , Chang Hua , Landesman Yosef , Charlson John , Pradeep Sunila
Published: June 2022

Leiomyosarcoma (LMS) is a rare soft tissue sarcoma (STS) that begins in smooth muscle tissue and most often initiates in the abdomen or uterus. Compared with other uterine cancers, uterine LMS is an aggressive tumor associated with poor prognosis and has a high risk of recurrence and death, regardless of stage at presentation. Selinexor is a first-in-class selective inhibitor of nuclear export (SINE) compound that binds covalently to exportin 1, thereby reactivating tumor suppressor proteins and downregulating the expression of oncogenes and DNA damage repair (DDR) proteins. Selinexor has demonstrated preclinical activity in various cancer types, including STS, and is currently in Phase I and II clinical trials for advanced cancers (Marretta et al. 2021). In this study, we evaluated the effects of Selinexor in combination with Dacarbazine, Gemcitabine, Doxorubicin, Pazopanib, Eribulin, and Trabectedin in LMS cell lines. The effects on cell viability were evaluated by treating the LMS cell lines SK-UT1 (p53-mut) and SK-UT1-B (p53 WT) with Selinexor and different chemotherapy combinations. Treatment with these drugs significantly decreased the cell proliferation and survival of SK-UT1 and SK-UT1-B cell lines tested in vitro. However, this effect was enhanced in SK-UT1 cells following treatment in the combination of Selinexor with Eribulin and Doxorubicin. The effects of treatment were also evaluated in mouse xenograft models generated with SK-UT1 cell lines. Treatment with Selinexor combined with Eribulin showed synergistic effects on tumor growth inhibition in SK-UT1 derived xenografts which warrant further investigation of these combinations at the clinical level.

Citation Format: Sonam Mittal, Ishaque Pulikkal, Hua Chang, Yosef Landesman, John Charlson, Sunila Pradeep. Selinexor, a selective inhibitor of nuclear export (SINE) compound, enhances the anti-tumor activity of Eribulin in leiomyosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5314.
Abstract 5325: Image-based functional precision medicine for repurposing neuroactive drugs in glioblastoma
Authors: Lee Sohyon , Weiss Tobias , Bühler Marcel , Wegmann Rebekka , Mena Julien , Bihl Michel , Goetze Sandra , van Drogen Audrey , Rushing Elisabeth J. , Wollscheid Bernd , Weller Michael , Snijder Berend
Published: June 2022

Treatment of glioblastoma multiforme (GBM), the most aggressive form of primary brain cancer, has essentially not advanced over the past few decades. Numerous challenges hinder the successful development of new therapies, including drug delivery across the blood-brain barrier (BBB), the complexity of the tumor microenvironment (TME), and the lack of clinically predictive cancer models. Here, we present the results of an image-based ex vivo drug-testing platform that addresses these therapeutic roadblocks. To demonstrate the clinical utility of our platform, in a retrospective cohort of 14 GBM patients, we show that ex vivo sensitivity to Temozolomide (TMZ, 1st-line GBM chemotherapy), is associated with longer progression free survival (PFS) and overall survival (OS). Next, by screening 150 clinically approved drugs across 27 GBM surgical patient samples, we identify a set of BBB-permeable neuroactive drugs with anti-glioma activity. These neurological drugs display remarkably consistent on-target killing of cancer cells with minimal toxicity to non-malignant TME cells across both primary and recurrent GBM samples. Single-cell transcriptional profiling of GBM patient samples and functional genetics reveals novel glioma-dependencies on neurological drug-target expression. Furthermore, a drug-target network enrichment analysis uncovers an AP1/BTG/TP53 gene signature associated with the anti-glioma activity of neurological drugs. In silico screening of over 1 million compounds for this common gene signature identified additional drug hits that could be validated in patient samples with 90% accuracy. Multiplexed transcriptomics revealed AP-1 transcription factor family activation to be the common underlying feature of neurological drugs with anti-glioma activity. Among the most promising candidate drugs, we identify the atypical antidepressant Vortioxetine as the strongest inducer of this gene signature, and confirm its efficacy in vivo across multiple mouse models. Vortioxetine in combination with Temozolomide or Lomustine further increased median survival in vivo compared to single agents alone. This study thus provides a clinically predictive and personalized drug-testing platform that identifies new treatment opportunities for GBM, warranting further investigation.

Citation Format: Sohyon Lee, Tobias Weiss, Marcel Bühler, Rebekka Wegmann, Julien Mena, Michel Bihl, Sandra Goetze, Audrey van Drogen, Elisabeth J. Rushing, Bernd Wollscheid, Michael Weller, Berend Snijder. Image-based functional precision medicine for repurposing neuroactive drugs in glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5325.
Abstract 533: A study of sequencing circulating cell free DNA from lung cancer patients
Authors: Kang Zhigang , Cao Liang
Published: June 2022

Precision oncology requires accurate cancer molecular information. While widely used to characterize tumors, tissue biopsy is limited in patient follow-up due to its invasive nature and the lack representation of the entire tumor bulk. To develop the method of circulating cell free DNA (cfDNA) for obtaining clinically valuable sequence information, we isolated cfDNA from the plasma samples of 10 advanced metastatic lung cancer patients and performed next generation sequencing (NGS) using a unique molecular marker (UMI) integrated 275-gene-target-panel at the average depth of 4,000 X coverage. We were able to identify common mutants of lung cancer driver genes, e.g. EGFR, KRAS, DNMT3 and TP53 with high confidence. In one example, the TKI resistant EGFR mutant T790M was found coexisting with EGFR 746-750del and TP53 splice variant in a baseline sample of a patient progressed from a first-generation EGFR inhibitor. And the dynamic changes of the allele frequency of EGFR and TP53 mutants were consistent with total cfDNA levels of the patients. Interestingly, an osimertinib resistance EGFR mutant C797S emerged at Day 355 post-treatment and followed by disease progression in 4 months. In addition to EGFR gene mutations, we further found increasing EGFR copy number associated with drug resistance and cancer progression of this patient. Our study demonstrated that it is feasible using cfDNA for patient follow-up to understand cancer resistance or disease progression through characterization of cancer mutations.

Citation Format: Zhigang Kang, Liang Cao. A study of sequencing circulating cell free DNA from lung cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 533.
Abstract 5358: Involvement of solute carrier transporters SLC25A17 and SLC27A6 in metabolic reprograming of castration-resistant prostate cancer
Authors: Kushwaha Prem P. , Verma Shiv S. , Shankar Eswar , Lin Spencer , Gupta Sanjay
Published: June 2022

Enzalutamide (XTANDI®), an antiandrogen, is used for the treatment of advance-stage prostatecancer. Approximately, 60% of patients receiving enzalutamide show initial remission followedby disease relapse with the emergence of highly aggressive castration-resistant prostate cancer.Solute carrier (SLC) transporters play a critical role in cancer drug resistance by altering cellularmetabolism. We have recently reported the predominance of SLC25A17 and SLC27A6 duringenzalutamide resistance (Cells 9(12):2535, 2020) by generating enzalutamide resistant (C4-2B-Enzu) cells exposing human prostate cancer C4-2B cells in the presence of increasingconcentration of enzalutamide (1-20 µM) and maintained in the same condition for six months.However, the role of these molecules during metabolic reprogramming have not beenelucidated. Knockdown of SLC25A17 and SLC27A6 gene by sgRNA transfection suppressedcell proliferation, and migration in C4-2B-Enzu cells. Analysis of cell cycle showed that in theabsence of SLC25A17 and SLC27A6 there is induction of G1/S cell cycle arrest and reductionin the protein expression of checkpoint factors viz. Cyclin D1, CDK6, CDK4, and CDK2.SLC25A17 and SLC27A6 knockdown resulted in downregulation of acetyl-CoA carboxylase(ACC), fatty acid synthase (FASN) and Acyl-CoA synthetase long chain family member 5(ACSL5) with simultaneous decrease in the levels of lactic acid, triglyceride and citric acid in C4-2B-Enzu cells. In addition, SLC25A17 knockdown resulted in marked increase in apoptosis asevident by increase in protein expression of RIPK3 and BAX, compared to SLC27A6 knockdownin C4-2B-Enzu cells. Taken together, our results demonstrate that solute carriers SLC25A17and SLC27A6 play an important role in the development of enzalutamide resistance throughmetabolic reprogramming and loss of these molecules results in cell death. Thus, SLC25A17and SLC27A6 may serve as a therapeutic target for enzalutamide resistant prostate cancer.

Citation Format: Prem P. Kushwaha, Shiv S. Verma, Eswar Shankar, Spencer Lin, Sanjay Gupta. Involvement of solute carrier transporters SLC25A17 and SLC27A6 in metabolic reprograming of castration-resistant prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5358.
Abstract 5371: Repurposing erbb2 (HER2/neu) as a neoantigen target for neu-negative tumors using a novel erbb2-encoding lentivirus
Authors: Gabriel Emmanuel M. , Necela Brian , Bahr Deborah , Kaplan Jamie , Attwood Kristopher , Knutson Keith
Published: June 2022

INTRODUCTION: HER2/neu is a potent biomarker for which anti-HER2/neu antibodies offer significant responses. However, many cancers, such as melanoma, have less than 5% expression of HER2/neu. The purpose of this study was to determine whether induced expression of erbb2 (mouse HER2/neu analogue) could function as a neoantigen target for anti-erbb2 antibody.

METHODS: RT-PCR was used to construct a rat erbb2-encoding vector using the live, non-replicating lentivirus pLenti6.3. erbb2-encoding pLenti6.3 was transfected in vitro into the B16 mouse melanoma cell line, which has less than 5% endogenous expression of erbb2. The generated erbb2+ B16 cell line was designated B16/neu. Anti-erbb2 antibody (7.16.4) was used to treat C57Bl/6 male and female mice bearing B16/neu (200 ug intraperitoneally three times per week for 3 weeks). To confirm activity of 7.16.4, this antibody (100 ug per T75 flask) was co-cultured with B16/neu in vitro, and cell growth/confluence was measured.

RESULTS: The novel B16/neu cell line expressed approximately 50% erbb2 in vitro on flow cytometry analysis, compared to 2% errb2 expression in naïve B16. Erbb2/neu expression was confirmed on fluorescent Western blot. Anti-erbb2 antibody (7.16.4) slowed the growth of B16/neu in vitro, confirming the suppressive effects of 7.16.4. In B16/neu-bearing mice, 7.16.4 resulted in effective control of tumor growth and a complete tumor response rate of 35.7%, (5/14 mice) compared to placebo controls (p < 0.01). Interestingly, B16/neu tumors demonstrated slower growth kinetics compared to naïve B16 tumors (p = 0.013), suggesting immune recognition of erbb2 in naïve C57Bl/6 mice as a neoantigen target. No adverse reactions were observed with 7.16.4. In vivo inoculation of erbb2-encoding pLenti6.3 in naïve B16 tumors and treatment of in vivo transfected B16/neu tumors with 7.16.4 comprise ongoing experiments.

CONCLUSIONS: erbb2 expression on B16 melanoma was successfully achieved using a novel erbb2-encoding lentivirus. Induced erbb2 served as a neoantigen target for anti-erbb2 antibody. This preclinical study provides translational evidence that induced HER2/neu expression may be utilized to repurpose anti-HER2 antibodies for human malignancies that do not normally express high levels of HER2/neu.

Citation Format: Emmanuel M. Gabriel, Brian Necela, Deborah Bahr, Jamie Kaplan, Kristopher Attwood, Keith Knutson. Repurposing erbb2 (HER2/neu) as a neoantigen target for neu-negative tumors using a novel erbb2-encoding lentivirus [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5371.