tbuckley/cabot-search huggingface dataset:
Abstract 4802: Functional analysis of cancer associated mitochondrial genome alterations
Authors: Neil Otto
Journal: Cancer Research (2020)

Abstract It is well documented that a variety of alterations can occur in the mitochondrial genome during human tumor development. These alterations often correlate with cancer phenotypes like treatment resistance and metastasis. Alterations to the mitochondrial genome may therefore be a biomarker for early detection, prognosis and/or predictor of therapeutic sensitivity. However, a method to easily and quickly create isogenic populations of human cells with mitochondrial genome specific alterations is not available. To combat this, we have developed a technology using transient expression of mitochondria targeted TAL domain nickases and TAL endonucleases (TALENs). This the first reliable method to introduce specific mutations into the mitochondrial genome of mammalian cells. We have efficiently induced mtDNA deletions in zebrafish embryos, human and mouse cell lines. In mouse NIH3T3 fibroblasts, we deleted a mitochondrial genome region corresponding to the same disease sequence seen in the human Kearns-Sayre Syndrome (KSS). The deletion resulted in a measurable phenotype including decreased mitochondria basal respiration, ATP production and maximal respiration. We are expanding this work by engineering targeted and precise mtDNA deletions in conjunction with known cancer initiating nuclear mutations in mammalian cells, as well as systemically in mice. These tools will allow us to determine the effect mitochondrial alterations have on the transformation of cancer cells harboring known cancer causing nuclear mutations. We are also developing an inducible system for expressing mitoTALENs, making it possible to create “conditional” loss of function mutations in mitochondrial genes in human cell lines, including induced pluripotent stem cells (iPSCs). This inducible model system will be highly useful in settings where the mitochondrial deletion may cause a selective disadvantage during early cellular differentiation. Citation Format: Neil Otto. Functional analysis of cancer associated mitochondrial genome alterations [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 4802.
Abstract 4438: A bispecific antibody targeting CD28H and PDL-1 is a novel and potent immunomodulator of T cell responses
Authors: Madhu Ramaswamy, Taeil Kim, Desmond Jones, Hormas Ghadially, Tamer Mahmoud, Andrew Garcia, Susan Wilson, Jeffrey Riggs, Darren Schofield, GIanluca Carlesso
Journal: Cancer Research (2020)

Abstract The success of immune checkpoint inhibitors CTLA4 and PD-1 monoclonal antibody, both FDA approved cancer therapies, specifically counteracts inhibitory pathways to activate antigen-specific T cells. However, their success is limited by the fact that not all patients respond to immunotherapy and a variety of adverse events due to non specific and systemic T cell activation limits their administration. A newly identified B7 family receptor, CD28H/TMIGD2, is constitutively expressed on T, pDCs and innate lymphoid cells including NK and ILCs. CD28H is also detected in TILs including the CD8+ tissue resident memory T cells (TRM), a T cell subset that correlates with better prognosis and response to immune checkpoint inhibitor therapy. Functionally, CD28H provides costimulatory function to T cells in the context of TCR signaling events during activation. We engineered a bispecific antibody (Bis mAb) targeting CD28H and PDL1 aimed to augment T cell costimulation and NK activation. The CD28H-PDL1 Bis mAb potentiates T cell proliferative and cytokine responses in antigen-specific human T cell assays and induces human NK -mediated redirected killing of PDL1+ tumor cells. In T cells, the mechanism of action of the Bis mAb requires intracellular signaling domain of the CD28H which downmodulates SHP phosphorylation upon CD3 activation. Intriguingly, we found that the Bis mAb increases induction of CD8 TRM cells in vitro. In human TILs, where CD28H is mostly expressed on CD8 TRM in tumors, the Bis mAb enables higher cytokine responses over PDL1 mAb alone. Given the recent relevance of TRM cells as an important pool of anti-tumor T cell immunity, the rationale for targeting this population via CD28H in the context of blocking PDL1 may prove to be a therapeutic tool for enhancing responses to checkpoint inhibitor therapy. Citation Format: Madhu Ramaswamy, Taeil Kim, Desmond Jones, Hormas Ghadially, Tamer Mahmoud, Andrew Garcia, Susan Wilson, Jeffrey Riggs, Darren Schofield, GIanluca Carlesso. A bispecific antibody targeting CD28H and PDL-1 is a novel and potent immunomodulator of T cell responses [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 4438.
プラスミド/ポリカチオン/RGD-PEG-COOH三元複合系を含む新しいレセプター媒介遺伝子トランスフェクション系
Authors: Sakae M, Ito T, Tanaka Ni, Niidome T, Yanagie H, Koyama Y
Journal: Molecular Therapy (2004)
What’s in a Number? Examining the Prognostic and Predictive Importance of Platelet Count in Patients With Essential Thrombocythemia
Authors: Andrew T. Kuykendall, Rami Komrokji
Journal: Journal of the National Comprehensive Cancer Network (2020)

Essential thrombocythemia (ET) is a myeloproliferative neoplasm characterized by clonal overproduction of platelets and an increased risk of thrombohemorrhagic complications. Patients are risk stratified by driver mutation, age, and thrombotic history and treated to reduce the risk of thrombotic and hemorrhagic events. The significance of platelet number as a risk factor or treatment goal is unclear. Despite the preponderance of data failing to demonstrate an association, there exists a pervasive belief that higher platelet counts correlate with an increased thrombotic risk. In fact, the association between thrombocytosis and bleeding is more clearly supported. Variability in regional consensus guidelines contributes to the uncertainty. This article reviews the data that shed light on the importance of platelet count in patients with ET.
Abstract 1134: Developing an early expression-based signature to assess interval colorectal carcinoma risk
Authors: Peh Yean Cheah, Lai Fun Thean, Kuen Kuen Lam, Soo Chin Liew, Michelle Wong, Nicholas Koh, Michelle Lo, Emile Tan, Choong Leong Tang
Journal: Cancer Research (2020)

Abstract Colorectal cancer (CRC) is the third most frequent cancer and the second leading cause of cancer mortality in the developed world including Singapore. Despite an aging populace which is now more compliant to having a baseline colonoscopy at age 50 or earlier, interval cancer (cancer that develops in between colonoscopies) is an unmet critical problem. The identification of early driver genes that can serve as biomarkers for CRC risk will aid in clinical management decision to counter cancer-associated mortality and morbidity. We profiled the genome-wide expression of matched mucosa (M), mucosa next to tumor (MT) and tumor (T) samples of a series of 23 patients using Affymetrix Gene Array. M samples are at least 20 cm away from the tumor and MT samples are adjoining the T samples but maintained the normal colonic crypt architecture of M. We discovered that thirty-nine dysregulated genes in MT samples compared to M samples persists until tumor formation. These early driver genes comprise not only tumor suppressors or oncogenes but included metabolic and transporter genes indicating that transformation towards malignancy requires the disruption of many pathways. Majority of these genes were down-regulated; the top three most drastically down-regulated genes include AQP8 (a water transporter), CA7 (a metabolic gene) and PHLPP2 (a tumor suppressor). Further analysis indicates that the three genes were already down-regulated in the normal appearing mucosa of a ‘healthy' individual compared to age-, gender-, ethnicity-matched (AGE-) controls seven years before recto-sigmoid cancer was diagnosed. This suggests that the dysregulation can occur several years before tumor formation and hence could possibly be used as early or pre-cancer biomarker. We have validated the down-regulation of the 3 genes in an independent series of 135 CRC samples from the same center using real-time RT-PCR assays. Correlation of the expression with clinicopathological features of the patients (gender, tumor site, age-group) and molecular subtypes (CMS 1-4) of the tumors will be ascertained to assess whether the signature is applicable to all CRC subtypes. Furthermore, the expression in matched blood samples will also be determined to explore feasibility of using these samples for assessing interval colorectal carcinoma risk. Citation Format: Peh Yean Cheah, Lai Fun Thean, Kuen Kuen Lam, Soo Chin Liew, Michelle Wong, Nicholas Koh, Michelle Lo, Emile Tan, Choong Leong Tang. Developing an early expression-based signature to assess interval colorectal carcinoma risk [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 1134.
Abstract 5267: Utilizing stimulated Raman scattering microscopy to study intracellular distribution of label-free ponatinib in live cells
Authors: Kristel Sepp, Martin Lee, Marie T. Bluntzer, G. Vignir Helgason, Alison N. Hulme, Valerie G. Brunton
Journal: Cancer Research (2020)

Abstract Ponatinib is a clinically approved tyrosine kinase inhibitor used to treat chronic myeloid leukemia (CML). Drug resistance is a widespread problem in CML treatment, where ponatinib resistant patients have very limited treatment options. In this study stimulated Raman scattering (SRS) microscopy was used to allow label-free imaging of intracellular ponatinib with high sensitivity and specificity in live human CML cell lines, in the context of ponatinib resistance. Ponatinib has an alkyne moiety in its structure that makes it inherently Raman active in the cellular silent region of the Raman spectrum. SRS microscopy represents a powerful imaging tool for visualizing label-free drug molecules within cells with high resolution, without the need for additional labels, or nanoparticle sensors as used in many other optical imaging technologies. It provides Raman imaging with minimal spectral distortion and a quantitative output, allowing the intracellular concentrations of drug molecules to be accurately determined. Intracellular visualization of ponatinib was achieved at biologically relevant, nanomolar concentrations for the first time using SRS. It was determined that ponatinib is sequestered into the lysosomes, with a higher lysosomal concentration found in drug resistant cells. This was associated with increased lysosome biogenesis. Target engagement studies showed that treatment with chloroquine reduced ponatinib accumulation in lysosomes, but did not re-sensitise cells to ponatinib, confirming BCR-ABL independent resistance mechanism in this CML cell model. In summary, we have visualized intracellular ponatinib localization for the first time using SRS, demonstrating that acquired drug resistance can influence drug uptake and localisation in CML, which in turn has an effect on target engagement. Citation Format: Kristel Sepp, Martin Lee, Marie T. Bluntzer, G. Vignir Helgason, Alison N. Hulme, Valerie G. Brunton. Utilizing stimulated Raman scattering microscopy to study intracellular distribution of label-free ponatinib in live 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 5267.
Functional and Biological Implications of CUX1 Mutations and Deletions in Myeloid Neoplasms
Authors: Mai Aly, Zubaidah M Ramdzan, Yasunobu Nagata, Suresh K. Balasubramanian, Naoko Hosono, Hideki Makishima, Valeria Visconte, Bartlomiej P. Przychodzen, Cassandra M. Hirsch, Mikkael A. Sekeres
Journal: Blood (2017)
Equivalent performance of single-use and reusable duodenoscopes in a randomised trial
Authors: Ji Young Bang, Robert Hawes, Shyam Varadarajulu
Journal: Gut (2020)

Single-use duodenoscopes have been recently developed to eliminate risk of infection transmission from contaminated reusable duodenoscopes. We compared performances of single-use and reusable duodenoscopes in patients undergoing endoscopic retrograde cholangiopancreatography (ERCP).Patients with native papilla requiring ERCP were randomised to single-use or reusable duodenoscope. Primary outcome was comparing number of attempts to achieve successful cannulation of desired duct. Secondary outcomes were technical performance that measured duodenoscope manoeuvrability, mechanical-imaging characteristics and ability to perform therapeutic interventions, need for advanced cannulation techniques or cross-over to alternate duodenoscope group to achieve ductal access and adverse events.98 patients were treated using single-use (n=48) or reusable (n=50) duodenoscopes with >80% graded as low-complexity procedures. While median number of attempts to achieve successful cannulation was significantly lower for single-use cohort (2 vs 5, p=0.013), ease of passage into stomach (p=0.047), image quality (p<0.001), image stability (p<0.001) and air-water button functionality (p<0.001) were significantly worse. There was no significant difference in rate of cannulation, adverse events including mortality (one patient in each group), need to cross-over or need for advanced cannulation techniques to achieve ductal access, between cohorts. On multivariate logistic regression analysis, only duodenoscope type (single-use) was associated with less than six attempts to achieve selective cannulation (p=0.012), when adjusted for patient demographics, procedural complexity and type of intervention.Given the overall safety profile and similar technical performance, single-use duodenoscopes represent an alternative to reusable duodenoscopes for performing low-complexity ERCP procedures in experienced hands.Clinicaltrials.gov number: NCT04143698.
Abstract 1816: β-catenin antagonist peptide attenuates oncogenic gene transactivation and promotes antitumor activity in breast cancer models
Authors: Jim A. Rotolo, Lila Ghamsari, Erin Gallagher, Rick Ramirez, Mark Koester, Siok Leong, Gene Merutka, Barry J. Kappel
Journal: Cancer Research (2020)

Abstract Aberrant activation of the canonical Wnt signaling pathway occurs in a variety of cancers, resulting in overactivation of β-catenin-mediated transcriptional activity and increased expression of genes that promote tumor survival, proliferation and inhibit differentiation. Disruption of the β-catenin transcription complex, by inhibiting protein-protein interactions required for its formation, represents a powerful approach to inhibit this previously ‘undruggable' target. The goal of these experiments is to characterize the anti-tumor activity of β-catenin antagonist peptides, BCA1 and BCA2, designed to disrupt the β-catenin transcription complex, in breast cancer models. In in vitro experiments, MCF7 (HER2neg/HRpos) breast adenocarcinoma cells were exposed to β-catenin antagonist peptides, and the impact on β-catenin-mediated gene transactivation was determined by quantitative polymerase chain reaction (qPCR). Results indicate that β-catenin antagonist peptide attenuated oncogenic gene transactivation in MCF7 cells, significantly decreasing the expression of direct β-catenin target genes including Myc, Cyclin D1 and Cyclin-dependent kinase 4 (p&amp;lt;0.05). The impact of β-catenin antagonist peptides on cancer cell viability was quantified by annexin V/PI flow cytometry analysis, indicating a dose-dependent decrease in overall cell viability 48 hours post peptide exposure. Additionally, BCA1 was administered to primary breast cancer tumoroids (HER2pos/HRneg or HER2neg/ HRneg), and the impact on tumoroid viability was quantified by histology and immunofluorescence microscopy. β-catenin antagonist peptide BCA1 resulted in dose-dependent decrease in surviving HER2pos/HRneg and HER2neg/ HRneg breast cancer tumoroids, with EC50 values of 5.2 and 6.9 µM, respectively. Finally, the impact of BCA2, an analogue of BCA1 with an additional N-terminal modification, on subcutaneous MCF7 xenograft tumors was determined in vivo. Administration of 5 mg/kg BCA2 via subcutaneous injection at a dosing frequency of 3x/week for 3 weeks resulted in 18-day tumor regression from the start of treatment and a 63.1% tumor growth inhibition compared to vehicle-treated controls (p&amp;lt;0.05). Taken together, these data demonstrate the significant anti-breast cancer activity of β-catenin antagonist peptides in vitro and in vivo. Citation Format: Jim A. Rotolo, Lila Ghamsari, Erin Gallagher, Rick Ramirez, Mark Koester, Siok Leong, Gene Merutka, Barry J. Kappel. β-catenin antagonist peptide attenuates oncogenic gene transactivation and promotes antitumor activity in breast cancer models [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 1816.
Abstract 4993: Label-free metabolic imaging of T cell response to the tumor microenvironment
Authors: Ava VanDommelen, Amani Gillette, Manish S. Patankar, Melissa Skala
Journal: Cancer Research (2020)

Abstract Numerous factors including low pH, low glucose, and high lactic acid levels in tumors create an immunosuppressive tumor microenvironment. This immunosuppression is partially due to the metabolic requirements of T cell activation, which are difficult to meet in the tumor microenvironment. The goal of this study is to use label-free optical metabolic imaging (OMI) of the metabolic co-factors NAD(P)H and FAD in CD3-conjugated (activated) and control T cells to monitor metabolic changes of T cells within tumor micro environmental conditions. Single-cell measurements include the optical redox ratio (fluorescence intensity of NAD(P)H divided by FAD), which measures the oxidation-reduction state of the cell, and the mean fluorescence lifetimes (τm) of NAD(P)H and FAD, which measure enzyme-binding activity in the cell. Jurkat T cells were conjugated with CD3 antibody over a two-day incubation period to achieve activation. Control and CD3-conjugated cells were exposed to environments with increasing concentrations of lactic acid (0-20 mM), hydrochloric acid (HCl; pH 6.4- 7.4), and glucose deprivation to mimic the tumor microenvironment. OMI monitored metabolic changes on a single-cell level without confounding factors associated with cell surface marker labeling. Treatment with lactic acid resulted in a significant dose-dependent decrease in the FAD τm and optical redox ratio (p&amp;lt;0.005). Along with changes in metabolism, OMI also resolved impaired mobility of the T cells with increasing lactic acid concentrations. Lactic acid affects both pH and metabolism, so cells were separately exposed to different concentrations of HCl (pH 6.4 - pH 7.4) to monitor pH effects alone. In control cells, the optical redox ratio decreased with decreasing pH (p&amp;lt;0.005). Conversely, in the presence of CD3, the optical redox ratio increased with decreasing pH (p&amp;lt;0.005). CD3 conjugation in glucose deprived environments also resulted in increases in both optical redox ratio and NAD(P)H τm. Cellular viability assays indicate apoptosis only in the lowest pH environment (pH 6.4, p&amp;lt;0.005). These studies indicate that OMI is a promising label-free approach to investigate the effects of the tumor microenvironment on T cell metabolism, activation, and drug response. Citation Format: Ava VanDommelen, Amani Gillette, Manish S. Patankar, Melissa Skala. Label-free metabolic imaging of T cell response to the tumor microenvironment [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 4993.
Abstract 3030: Different pharmacodynamic profiles of ERK1/2 inhibition can elicit comparable anti-tumor activity
Authors: Christopher Hindley, Luke Bevan, Hannah Braithwaite, Aurélie Courtin, Lynsey Fazal, Roberta Ferraldeschi, Vanessa Martins, Sandra Muench, Nicola Wallis, Nicola Wilsher, John Lyons, Joanne Munck
Journal: Cancer Research (2020)

Abstract The MAPK pathway is frequently dysregulated in cancer, resulting in constitutive phosphorylation and activation of its downstream signaling node ERK1/2 (ERK). The direct targeting of activated ERK is being explored in several phase I clinical trials. A good understanding of the relationship between inhibition of pathway activity and inhibition of tumor growth allows dose scheduling in the clinic to maximise response to an inhibitor. We have recently described the development of a novel, potent and selective small molecule inhibitor of both ERK activity and phosphorylation using fragment-based drug discovery. Using this inhibitor as a tool compound, we explored the relationship between duration of ERK inhibition and inhibition of tumor growth. We examined the in vivo activity of our ERK inhibitor in a Colo205 xenograft model. Inhibition of the ERK pathway was monitored by measuring phosphorylation of a direct substrate of ERK, RSK90 (pRSK), and phosphorylated ERK itself (pERK) to determine pharmacodynamic profiles. We observed that pathway inhibition was consistent with the pharmacokinetic profile of the ERK inhibitor and that the duration of pathway inhibition was primarily determined by dosing frequency rather than the total dose of ERK inhibitor. High, single doses (50 or 40 mg/kg) of ERK inhibitor produce strong maximal pathway inhibition at 2 h post dose (pRSK levels 8% of control levels for a single dose of 40 mg/kg) but pRSK levels have returned to levels close to control values by 12 h post dose. In contrast, 3x equally spaced doses of 20 mg/kg have a lower maximal pathway inhibition at 2 h post dose (pRSK levels 29% of control) but extend pathway inhibition beyond 12 h (pRSK levels 21% of control at 14 h). Despite these two different pharmacodynamic profiles, we observed comparable tumor growth inhibition following daily dosing either at 1 × 50 mg/kg (maximal tumor growth inhibition of 21%) or 3 × 20 mg/kg (maximal tumor growth inhibition of 15%). These data suggest that a short duration of ERK inhibition rather than constant suppression can be sufficient to provide anti-tumor activity. This may allow flexibility in clinical dosing schedules to improve tolerability without a loss in efficacy. Citation Format: Christopher Hindley, Luke Bevan, Hannah Braithwaite, Aurélie Courtin, Lynsey Fazal, Roberta Ferraldeschi, Vanessa Martins, Sandra Muench, Nicola Wallis, Nicola Wilsher, John Lyons, Joanne Munck. Different pharmacodynamic profiles of ERK1/2 inhibition can elicit comparable anti-tumor activity [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 3030.