bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract A49: Specific Kras codon 12 and 13 mutations display different tumor initiation in pancreatic cancer
Authors: Martin Maria Paz Zafra , Alonso-Curbelo Direna , Wilkinson J. Erby , Schatoff Emma , Dow Lukas E. 
Published: June 2021

Abstract
Defining the effect of specific oncogenic mutations on tumor behavior and therapy response is essential for developing effective treatment strategies and realizing the promise of precision medicine. KRAS is the most frequently mutated oncogene in human cancers, and specific cancer types show a clear bias in the types and frequency of Kras alterations. The generation of conditional animal models, such as Lox-Stop-Lox (LSL) KrasG12D (KC model) and LSL-KrasG12Vgeo mice, has provided important tools to dissect the impact of Kras mutation in tumor development, but these models alone do not recapitulate the spectrum of Kras alterations in human cancer. To better reflect this Kras mutational landscape, we have developed a series of critical LSL-Kras mouse models that recapitulate common Kras alterations observed in colorectal (G13D), pancreatic (G12R), and lung cancer (G12C). Mice carrying each LSL-Kras variant, a pancreas-specific Cre (p48-Cre) and a far-red fluorescent Cre-reporter (mKate2), displayed drastically different morphologic features in early stages of pancreatic cell transformation, associated with dramatic changes in the degree of stromal expansion within the pancreas. Now, we are focused in exploring how such changes following Kras alterations influence progression to carcinoma and response to targeted therapies.
Citation Format: Maria Paz Zafra Martin, Direna Alonso-Curbelo, J. Erby Wilkinson, Emma Schatoff, Lukas E. Dow. Specific Kras codon 12 and 13 mutations display different tumor initiation in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr A49.
Abstract B09: Combining proteomics and genetic screens to identify KRAS synthetic lethal interactions
Authors: Sheth Shikha S. , Cook Danielle R. , Strasser Samantha D. , Martin Timothy D. , Menon Sneha , Popow Olesja , Paulo Joao A. , Elledge Steve J. , Haigis Kevin M. 
Published: June 2021

Abstract
KRAS is a member of the RAS superfamily of small GTPases, proteins that act as molecular switches regulating cellular pathways such as growth, survival, and membrane trafficking. Activating point mutations in KRAS are found in approximately 15% of all cancers and contribute to a malignant phenotype through deregulation of normal cell proliferation. Despite discovering RAS mutations in cancer over three decades ago and strong evidence that RAS mutations drive cancer progression and response to therapy, there have been limited advances in targeting RAS directly. In lieu of direct inhibitors, identifying synthetic lethal (SL) partners with mutant KRAS would reveal putative drug targets. This strategy has been employed previously but with limited success. We hypothesize that using tools such as novel conditional mutant KRAS isogenic cell lines and CRISPR-based loss-of-function screening will uncover pathways whose depletion selectively impairs mutant KRAS cells. Additionally, approaching this from a multi-omics angle, where we combine proteomics and CRISPR screening, will allow us to prioritize targets from each screen. We used CRISPR/Cas9-mediated genome engineering to generate a KRAS WT colon cancer cell line with conditional expression of mutant KRAS. Characterization of our isogenic cell lines confirms that KRAS G12D expression upregulates KRAS activity and enhances tumorigenicity. We used a CRISPR library targeting druggable genes and screened these isogenic cells in vitro, and additionally performed mass spectrometry on the isogenic pair to look at the differential regulation of proteins and phospho-proteins upon KRAS G12D expression. We plan to determine if the overlap of these datasets can provide us with a prioritized list of hits to validate, arguing for a multi-omics approach to better understand synthetic lethal interactions.
Citation Format: Shikha S. Sheth, Danielle R. Cook, Samantha D. Strasser, Timothy D. Martin, Sneha Menon, Olesja Popow, Joao A. Paulo, Steve J. Elledge, Kevin M. Haigis. Combining proteomics and genetic screens to identify KRAS synthetic lethal interactions [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr B09.
Abstract B29: SHANK3 in oncogenic RAS signaling
Authors: Lilja Johanna , Jcaquement Guillaume , Posada Itziar Martinez D. , Laasola Petra , Vesilahti Ella-Maria , Miihkinen Mitro , Salomaa Siiri , Kreienkamp Hans-Juergen , Barsukov Igor , Abankwa Daniel , Ivaska Johanna 
Published: June 2021

Abstract
RAS proteins play a direct causal role in human cancers, with activating mutations in RAS occurring in ∼30% of tumors. Despite comprehensive efforts, efficient targeting of RAS has not yet reached the clinic. Here, we report SHANK3, a synaptic scaffolding protein and a regulator of integrin activity, as a novel tumor suppressor targeting RAS proto-oncogenes, especially mutant KRAS. We previously found the N-terminal part of SHANK3 as an unexpected RAS-association domain with high affinity for active RAS proteins. Now we show that the direct binding of SHANK3 to active RAS abrogates the MAPK/ERK signaling and thereby inhibits the RAS-stimulated transformation and slows the growth of RAS-dependent tumor cells. SHANK3 expression is downregulated in RAS-driven human cancers, and high expression of SHANK3 correlates with improved patient survival in KRAS-driven pancreatic and lung adenocarcinoma. These data support that targeting the MAPK/ERK pathway by SHANK3 represents a novel promising therapeutic approach for RAS-driven cancers.
Citation Format: Johanna Lilja, Guillaume Jcaquement, Itziar Martinez D. Posada, Petra Laasola, Ella-Maria Vesilahti, Mitro Miihkinen, Siiri Salomaa, Hans-Juergen Kreienkamp, Igor Barsukov, Daniel Abankwa, Johanna Ivaska. SHANK3 in oncogenic RAS signaling [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr B29.
Abstract B36: Berberine induces apoptosis in cervical carcinoma cells by inducing DNA damage and inhibition of RAS MAPK pathway
Authors: Komal Komal , Athreyas Vidhya , Chaudhary Shilpi , Singh Mayank 
Published: June 2021

Abstract
Cervical cancer remains the leading cause of cancer mortality in females worldwide. Despite significant improvement in treatment modalities, the survival in advanced cervical cancer is still limited, resulting in poor survival. Berberine is an isoquinoline alkaloid isolated from Berberis vulgaris (barberry). Berberine is in focus for its preventive and therapeutic value in different cancers. Here we investigated how berberine modulated DNA damage response and survival pathway in cervical carcinoma cells. We carried out the current study in both HPV-positive and -negative cervical carcinoma cell lines. Our results indicate that berberine induced apoptosis in all the cervical carcinoma cells through activation of both caspase-dependent and -independent pathways. Berberine also induces accumulation of gamma H2AX foci in cervical carcinoma cells, which colocalized with proteins involved in DNA damage response, indicating accumulation of replication stress in berberine-treated cells. Berberine also repressed the proliferative RAS MAPK pathway, thereby reducing the proliferation of cervical carcinoma cells. Overall our data provide a novel insight into mechanism of berberine action in cervical carcinoma cells and underline its efficacy as a chemoadjuvant in treatment of cervical cancer.
Citation Format: Komal Komal, Vidhya Athreyas, Shilpi Chaudhary, Mayank Singh. Berberine induces apoptosis in cervical carcinoma cells by inducing DNA damage and inhibition of RAS MAPK pathway [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr B36.
Abstract IA09: Targeting K-RAS4B on biologic membranes
Authors: Ikura Mitsu 
Published: June 2021

Abstract
RAS proteins are frequently mutated in cancer (~30% of all human tumors) and an estimated worldwide death toll of RAS-associated cancers exceeds 2 million/yr. Despite enormous efforts in RAS research over three decades, there is no clinically approved RAS inhibitor and the RAS protein remains a challenging target for cancer therapy development. In order to overcome this challenge, we sought to better understand how RAS functions under physiologic conditions and alters related signaling pathways in the mutant RAS-driven tumors. Fully matured protein K-RAS4B, the major target for cancer therapeutics, is prenylated and methylated at the carboxy-terminus, which enables K-RAS4B to anchor to the plasma membrane where it receives an upstream signal and transmits the signal to a number of downsteam pathways. There is, however, a large gap in our understanding of how the matured K-RAS4B protein functions at the surface of the plasma membrane. As a first step, we sought to develop new conformational and functional assays for the RAS protein on lipid bilayers (i.e., nanodiscs) using various biochemical and biophysical techniques. We elucidated how the membrane environment dictates the conformation of K-RAS4B and how oncogenic mutations influence the membrane-dependent conformational states of the protein (Mazhab-Jafari et al., PNAS 2015). More recently, we have been investigating multiple aspects of K-RAS4B functions and I will discuss (i) how the biologic membrane influences K-RAS4B interaction with a binding domain of RAF kinases and (ii) how we could inhibit K-RAS4B at the membrane surface by small molecules (Fang et al., Cell Chem Biol 2018) and an engineered protein. Supported by CCS, CIHR, CFI and PMCF.
Citation Format: Mitsu Ikura. Targeting K-RAS4B on biologic membranes [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr IA09.
Abstract IA12: Targeting KRAS-regulated metabolic activities in pancreatic cancer
Authors: Bryant Kirsten L. , Hobbs G. Aaron , Der Channing J. 
Published: June 2021

Abstract
One promising direction for the development of therapeutic strategies for the treatment of KRAS-driven pancreatic ductal adenocarcinoma (PDAC) growth involves delineating the role of and mechanism for altered metabolism. KRAS-mutant PDAC is characterized by increased autophagy and macropinocytosis, processes that enhance nutrient availability that then supports the increased metabolic needs of PDAC. First, we addressed the role of KRAS in supporting autophagy-dependent PDAC growth. Surprisingly, we found that suppression of KRAS increased autophagic flux, as did pharmacologic inhibition of its effector ERK MAPK. We speculated that ERK inhibition might enhance PDAC dependence on autophagy by impairing KRAS-driven metabolic processes, and we demonstrate that either KRAS suppression or ERK inhibition decreased both glycolytic and mitochondrial functions. Furthermore, genetic or pharmacologic inhibition of autophagy synergistically enhanced ERK inhibitor-mediated antitumor activity. We conclude that concurrent inhibition of ERK and autophagy may be an effective PDAC treatment. Second, we evaluated mechanisms by which KRAS may drive macropinocytosis. These studies focused on KRAS G12R mutant PDAC. This mutation is rare in lung and colorectal cancers (~1%), yet high (~20%) in PDAC. We evaluated whether KRAS G12R is functionally distinct from the more common KRAS G12D/V mutant proteins. We found that KRAS G12D/V but not KRASG12R drives elevated macropinocytosis and that MYC is essential for this process in KRAS G12D/V but not KRAS G12R mutant PDAC. The KRAS G12R mutation uniquely disrupts interaction with a key effector, PI3K alpha, due to structural perturbations in the switch II region critical for effector interaction. Despite this defect, KRAS G12R-mutant PDAC displayed elevated macropinocytosis due to KRAS-independent PI3K gamma activity. Finally, the growth of KRAS G12R mutant PDAC in vitro and in vivo was preferentially sensitive to MEK-ERK inhibition compared to KRAS G12D/V. In summary, our elucidation of KRAS-dependent effector signaling mechanisms that drive altered metabolism identified novel therapeutic approaches for the treatment of PDAC.
Citation Format: Kirsten L. Bryant, G. Aaron Hobbs, Channing J. Der. Targeting KRAS-regulated metabolic activities in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr IA12.
Abstract PR01: Novel state I structures of oncogenic KRAS4b mutants bound to GTP analog
Authors: Chan Albert H. , Tran Timothy H. , Dharmaiah Srisathiyanarayanan , Nissley Dwight V. , Simanshu Dhirendra K. 
Published: June 2021

Abstract
The RAS family proteins alternate between active GTP-bound and inactive GDP-bound states. Due to the high affinity of GDP and GTP, and low intrinsic GTPase activity of RAS, switching between the two states is assisted by guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). Many of the oncogenic RAS mutations result in impaired intrinsic and/or GAP-mediated GTPase function, leading to accumulation of constitutively active RAS. To understand the molecular mechanism behind the impairment, we crystallized and solved the structures of wild-type and six different KRAS4b oncogenic mutants bound to a non-hydrolysable GTP analog GMPPNP. Interestingly, we noticed variations in the conformation of the Switch I region, which contacts the nucleotide and many downstream effector proteins. In three of our mutants (G12C, G13D, Q61L), Switch I adopts an open conformation known as state I, which, similar to the GDP-bound state, does not contact the nucleotide and represents an inactive state of the RAS protein. Importantly, our state I structures revealed a new pocket created by the open Switch I loop that could be targeted in drug discovery. To screen for compounds that bind to this new Switch I pocket or other places of the protein, we started a fragment-screening campaign by X-ray crystallography using the KRAS4b-G13D-GMPPNP crystals. These crystals consistently diffracted to very high (~1.2 Å) resolutions, can be reproduced easily, are tolerant to DMSO soaking, and their putative fragment-binding pockets are unobstructed by lattice contacts, making them ideal for fragment-screening.
This abstract is also being presented as Poster A01.
Citation Format: Albert H. Chan, Timothy H. Tran, Srisathiyanarayanan Dharmaiah, Dwight V. Nissley, Dhirendra K. Simanshu. Novel state I structures of oncogenic KRAS4b mutants bound to GTP analog [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr PR01.
Abstract A30: Molecular insight into pathogen-associated molecular pattern signaling during TNFa and IL2 armed oncolytic adenovirus treatments
Authors: Heiniö Camilla , Havunen Riikka , Siurala Mikko , Hemminki Akseli 
Published: June 2021

Abstract
Adenoviral-mediated gene therapy is a promising new treatment for malignant tumors with poor responses to traditional cancer treatments. In this study, we investigate the effects of danger- and pathogen-associated molecular pattern (DAMP and PAMP) signaling responses to a chimeric, cytokine armed, oncolytic adenovirus (Ad5/3-E2F-d24-hTNFα-IRES-hIL2). It has previously been shown that this virus has a two-part mode of action against tumors. It simultaneously acts as an immunostimulatory agent while the virus itself lyses cancer cells. However, as this reaction is not fully understood on a molecular level, more in-depth information on the adenovirus-induced signaling cascades in the tumor microenvironment is needed. There are several pattern recognition receptors (PRR) in cells that are responsible for inducing the signaling cascades and consequent immune response towards DAMPs and PAMPs present during adenovirus infections. It has previously been reported that the most important PRRs and sensors include TLR9, NLRP3, RIG-1, cGAS, and AIM2. Binding of DAMPs and PAMPs to PRRs results in the activation of signaling cascades consisting of a plethora of proteins, finally culminating in the production and release of cytokines, pyroptosis or cellular activation and extracellular signaling, consequently affecting neighboring cells as well. In this study, several methods such as Western blot, flow cytometry and RNA-sequencing were used to study DAMP and PAMP signaling and the subsequent reactions in the tumor microenvironment. The knowledge gained about the signaling pathways in this study provides new insight into cell signaling, which can be used to further develop immuno- and virotherapy and to provide a new method of treating poorly responsive tumors.
Citation Format: Camilla Heiniö, Riikka Havunen, Mikko Siurala, Akseli Hemminki. Molecular insight into pathogen-associated molecular pattern signaling during TNFa and IL2 armed oncolytic adenovirus treatments [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A30.
Abstract B73: Development of anti-human CLDN18.2 monoclonal antibody as cancer therapeutics
Authors: Lin Haishan , Zhang Richard 
Published: June 2021

Abstract
Claudin-18 (CLDN18) is a member of a large family of four-span transmembrane proteins, claudins. These proteins are the essential components of the mammalian tight junctions (TJs) in the epithelial cells. Claudin-18 has two splice variants, 18.1 and 18.2. While CLDN18.1 is specifically expressed in the lung tissue, CLDN18.2 expression in normal tissue is more restricted and is only detected in small patches of stomach mucosal. CLDN18.2 expression is elevated in many types of epithelial cancers including stomach, esophagus, pancreatic and ovarian cancers. The expression of CLDN18.2 is not only detected in primary tumors, but also in the metastatic sites. Therefore, CLDN18.2 is an ideal target for monoclonal antibody-based cancer therapies. Anti-CLDN18.2 monoclonal antibody claudiximab (IMAB362) demonstrated promising results in the phase II clinic trial for gastric cancer. CLDN18.2 specific antibodies are difficult to generate due to its high homology to CLDN18.1. There are only 7-8 amino acid differences between these two splice variants in the first extracellular loop. Using a combination of DNA, protein, and transfected cells as immunogens, coupled with high-throughput FACS-based hybridoma screening approach, we have generated a panel of mouse monoclonal antibodies that are highly specific for CLDN 18.2. These antibodies had high affinities for CLDN18.2 while they did not bind to CLDN18.1. The lead antibodies had similar or better binding affinity for CLDN18.2, compared to the IMAB362. The anti-CLDN18.2 antibodies bound to human gastric carcinoma cell line KATO III. Our IHC data showed that the antibody specifically bound to gastric and pancreatic cancer tissues. In addition, we demonstrated that anti-CLDN18.2 antibodies induced receptor internalization in CHO cell line expressing CLDN18.2. The humanized anti-CLDN18.2 antibodies have been generated and showed similar binding specificity as the mouse antibodies. Lead anti-CLDN18.2 antibodies have been selected for CMC optimization and preclinical studies for further development as cancer therapeutics.
Citation Format: Haishan Lin, Richard Zhang. Development of anti-human CLDN18.2 monoclonal antibody as cancer therapeutics [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr B73.
Carotid Plaques From Symptomatic Patients Are Characterized by Local Increase in Xanthine Oxidase Expression
Authors: Ganji Morsaleh , Nardi Valentina , Prasad Megha , Jordan Kyra L. , Bois Melanie C. , Franchi Federico , Zhu Xiang Y. , Tang Hui , Young Melissa D. , Lerman Lilach O. , Lerman Amir 
Published: June 2021

Background and Purpose:
XO (xanthine oxidase) is a key enzyme of uric acid metabolism and is thought to contribute to oxidative pathways that promote atherosclerotic plaque progression, yet its role in plaque destabilization is not well elucidated. We hypothesized that XO is expressed in carotid plaque from symptomatic patients in association with cardiovascular risk factors.

Methods:
Patients were stratified by symptoms, defined as presentation with an ipsilateral cerebral ischemic event. Carotid atherosclerotic plaques were obtained from 44 patients with symptomatic plaque and 44 patients without ischemic cerebral events. Protein expression of XO was evaluated by immunohistochemical staining and the percentage of cells expressing XO and CD68 (macrophage marker) compared between the groups. Biochemical and demographic cardiometabolic risk factors of study participants also were measured.

Results:

Carotid atherosclerotic plaques from symptomatic patients were associated with significantly higher XO expression versus asymptomatic plaque (median [interquartile range]: 1.24 [2.09] versus 0.16 [0.34];
P
<0.001) and with significantly higher circulating uric acid levels (mean±SD: 7.36±2.10 versus 5.37±1.79 mg/dL;
P
<0.001, respectively). In addition, XO expression in atherosclerotic carotid plaque was inversely associated with serum high-density lipoproteins cholesterol levels (
P
=0.010,
r
=−0.30) and directly with circulating uric acid levels (
P
<0.001,
r
=0.45). The average percentage of macrophages that expressed XO was significantly higher in symptomatic versus asymptomatic plaques (median [interquartile range]: 93.37% [25] versus 46.15% [21], respectively;
P
<0.001).

Conclusions:
XO overexpression in macrophages is associated with increased serum uric acid and low high-density lipoproteins cholesterol levels and may potentially have a mechanistic role in carotid plaque destabilization. The current study supports a potential role for uric acid synthesis pathway as a target for management of carotid atherosclerosis in humans.
Parasites differentially impact crayfish personality in different contexts
Authors: MacKay Rebecca Noel , Moore Paul A. 
Published: June 2021

Abstract
The expression of an individual animal’s behaviour can be placed along many different personality spectra. Parasite load can alter animal behaviour and, thus, fitness. The personality traits of rusty crayfish, Faxonius rusticus, were analysed in three different behavioural contexts: foraging, exploration, and threatened. Each crayfish was tested in each context 3 times, giving a total of 9 assays per crayfish. After assays were completed, crayfish were dissected, and the hepatopancreas of each crayfish was photo analysed to determine the parasite load of the trematode, Microphallus spp. A composite personality score for each assay and parasite load was loaded into a PCA. The PCA model showed that as parasite load increased, crayfish became bolder in threatening contexts and less exploratory in novel environments, whether or not a food stimulus was present. Thus, parasite load alters the placement of crayfish on different personality spectra, but this change is context specific.