Pancreatic Cancer Cell Lines Research Articles

Abstract LB397: Simultaneous CRISPR-Cas9 induced double strand breaks causes chromosomal catastrophe in pancreatic cancers: A parallel comparison with ionizing radiation

Abstract CRISPR-Cas9, a highly effective gene editing tool, is a molecular scissor that induces double strand breaks (DSBs) at its targeted genomic locations. Ionizing radiation (IR), in contrast, generates DSBs and other DNA lesions, making it potentially more toxic than CRISPR-Cas9 induced DSBs. Alternatively, CRISPR-Cas9 might be more toxic as the Cas9 activity would continue if the DSB were repaired to wild type, resulting in more scissions per target. We designed multitarget sgRNAs targeting 0-16 sites in the human genome and performed similar dose-response experiments with IR (0-10 Gy) over 21 days on two pancreatic cancer (PC) cell lines. Growth inhibition was measured through clonogenicity and alamarBlue assays, and γ-H2AX staining performed to calculate DSBs. Surviving colonies from sgRNA transduction underwent whole genome sequencing (WGS) and targeted deep sequencing to quantify mutations. Cell growth was assessed through cell counting and sgRNA tag survival over time. Chromosomal changes were elucidated through chromosome breakage assays and XY FISH. We found that cytotoxicity increased as a function of both IR dose and number of sgRNA target site. At a comparable number of DSBs, CRISPR-Cas9 demonstrated higher cytotoxicity (~44% growth inhibition from 5-target sgRNA, ~15 γ-H2AX foci in the PC cells) compared to IR (9% inhibition from 0.5 Gy, ~25 foci). WGS of 5-cutter treated cells also revealed ~13 CRISPR-Cas9 induced DSBs in the PC cells. >99% cytotoxicity was achieved with 12-target sgRNA and 8 Gy. Moreover, sequencing of surviving colonies from sgRNA-treated cells confirmed lack of widespread off-target events. Notably, no surviving colonies could be obtained from the 12- and 16-cutter treated Panc10.05 cells and 8-14 cutters treated TS0111 cells.Both CRISPR-Cas9 and IR assays demonstrated delayed cell death, and cell death began 7 days after transduction of multitarget sgRNAs. Cytogenetic analyses on cells transduced with a 14-cutter sgRNA over 21 days revealed accumulation of karyotypic abnormalities peaking at day 14, and polyploidization peaking at day 10 and decreasing by day 21. Structural variant (SV) analyses from WGS data showed that the number of SVs per colony increased and peaked in 6-cutter colonies (average = 42 SVs), then decreased in 8- and 14-cutter colonies. Finally, surviving colonies that were resistant to 5-, 8-, and 16-cutter sgRNAs were re-challenged with their original multitarget sgRNA, 12-, and 14-cutter sgRNAs. Re-treating with original sgRNA showed no growth inhibition, while >95% cytotoxicity was observed by re-challenging them with 12- or 14-cutter sgRNAs.In conclusion, CRISPR-Cas9 kills pancreatic cancers by inducing chromosomal catastrophe, and resistance to original set of CRISPR-Cas9 induced DSBs can be easily overcome by targeting different sites. For the same number of DSBs, CRISPR-Cas9 was surprisingly more toxic than IR. Citation Format: Selina Shiqing K. Teh, Akhil Kotwal, Eitan Halper-Stromberg, Laura Morsberger, Kirsten Bowland, Alexis Bennett, Michael Goldstein, Ying S. Zou, James R. Eshleman. Simultaneous CRISPR-Cas9 induced double strand breaks causes chromosomal catastrophe in pancreatic cancers: A parallel comparison with ionizing radiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB397.

Abstract LB203: CRISPR anchor screens to identify co-vulnerabilities and drug combinations in DepMap

Abstract The Cancer Dependency Map (DepMap) aims to systematically map the landscape of cancer vulnerabilities by profiling a wide range of cancer cell lines that collectively represent the heterogeneity of human cancer. An increasing number of targets and biomarkers emerging from this effort are in drug development or clinical trials. However, it is clear that drug resistance is a significant challenge and that drug combinations will be required. CRISPR anchor screens involve introducing an "anchor" which can be either a drug or genetic perturbation, alongside the sgRNA library. This approach provides a way to investigate resistance mechanisms, potential drug combinations, or co-vulnerabilities. As a proof of concept, here we describe CRISPR anchor screens with MRTX1133 as the anchor, which is a potent and selective KRASG12D inhibitor currently in clinical trials. We performed genome-scale loss-of-function CRISPR-Cas12a screens in the presence of MRTX1133 in pancreatic and lung cancer cell lines that harbor the KRASG12D. The goal of these screens was to identify cancer dependencies that are either present or enhanced in the presence of MRTX1133. We find multiple such MRTX1133-induced dependencies that are either common across all cell lines screened or unique to one or a few. This shows that the response to MRTX1133 can vary depending on the context, highlighting the importance of finding biomarkers to predict the response. Identifying the biomarkers of these context-specific dependencies would provide strategies to select patient populations that would respond to the drug combinations. This study not only provides valuable insights into enhancing the efficacy of KRAS-targeted therapies but also serves as a pioneering effort in incorporating innovative CRISPR screen modalities into the DepMap. Citation Format: Jenny Serra-Vinardell, Zoe Posner, Alison Cameron, Yuqi Chen, Negin Karisani, Barbara De Kegel, Joshua M. Dempster, Andrew J. Aguirre, Francisca Vazquez. CRISPR anchor screens to identify co-vulnerabilities and drug combinations in DepMap [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB203.

Abstract LB192: AGR2 knockdown reduces cell proliferation and increases endoplasmic reticulum proteotoxic stress in pancreatic cancer

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal solid malignancy with low survival rates due to a dense stroma, abnormal gene expression patterns, and limited treatment options. Anterior gradient 2 (AGR2) is often overexpressed in many cancers, including PDAC. AGR2 belongs to the protein disulfide isomerase family of proteins and is located in the endoplasmic reticulum, where it helps with protein-disulfide isomerase reactions to ensure proper protein folding. However, the molecular mechanisms underpinning AGR2 overexpression in PDAC pathophysiology are not well understood. Gene expression data obtained from the pancreatic adenocarcinoma (PAAD) dataset shows that AGR2 is overexpressed in PDAC tumors compared to normal tissues. We confirmed this gene over-expression in PDAC tumors and multiple pancreatic cancer cell lines using fluorescent microscopy, RT-qPCR, and western blotting. Using stable inducible short hairpin (shRNA) technology, we showed that AGR2 knockdown significantly decreases the viability and colony-forming ability of HPAF-II and PANC-1 cells. Gene Ontology (GO) enrichment analysis revealed that AGR2 knockdown suppresses a host of protein biosynthesis pathways, including protein targeting and localization to the endoplasmic reticulum (ER). Significantly, ER stress was induced in knockdown cells, shown by the activation of unfolded protein response, suggesting an accumulation of unfolded proteins. To determine the effects of unfolded protein accumulation on the ER, we performed transmission electron microscopy. Here, the results show an increase in the ER lumen size of knockdown cells compared to controls. Interestingly, we observed a decrease in mitochondria size and confirmed the increased expression of mitochondria fission proteins DRP1 and MFF. Finally, flow cytometry analysis revealed increased apoptosis in AGR2 knockdown cells compared to controls. Overall, our data provide insights into the role of AGR2 in PDAC tumorigenesis. However, more research is warranted to determine if AGR2 could be a potential therapeutic target. Citation Format: Philip Salu, Katie Reindl. AGR2 knockdown reduces cell proliferation and increases endoplasmic reticulum proteotoxic stress in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB192.

Abstract LB274: Nuclear transporter HIKESHI is a novel independent poor overall survival for pancreatic cancer; targeting can inhibit pancreatic cancer growth via HSP70 suppression

Abstract HIKESHI, under heat stress conditions, has been reported to transport molecular chaperone HSP70 into the nucleus to prevent cancer cell death from heat stress. The tumor-supporting roles of molecular chaperones in various solid tumors are explored extensively. Yet, the sole targeting of the molecular chaperones could not significantly impact anticancer treatment. Thus, we hypothesized that the specific transporter protein HIKESHI might improve this outcome. However, understanding of the clinical relevance of HIKESHI in pancreatic cancer remained unclear, and few studies addressed the HIKESHI role in cancer and whether targeting can inhibit pancreatic cancer cell viability and the other HSPs. Here, we first investigated the expression pattern of the HIKESHI protein in surgically resected pancreatic cancer samples (n=142) and analyzed the clinical relevance of the HIKESHI. We found that the HIKESHI protein was overexpressed in cancer compared to the adjacent normal tissue, and positive expression of cancer HIKESHI was related to poor overall survival. In addition, HIKESHI-positive expression in cancer was identified as an independent prognostic marker for overall survival. Next, using shRNA, we established the stable HIKESHI-suppressed pancreatic cancer cell line (PANC-1). HIKESHI suppression inhibited cell growth and colony formation ability in vitro. The expression and nuclear translocation of HSP70 and HSP110/105 were suppressed in HIKESHI-suppressed cells compared to the control shRNA cells despite no heat stress conditions. Furthermore, we found that the tumor growth, HIKESHI, and Ki67 expression in HIKESHI-suppressed tumors were inhibited compared to the control shRNA tumors. Next, we performed Cap Analysis of Gene Expression and IP-MS analysis to discover the potential downstream and novel interactive partner of the HIKESHI protein. The extensive mechanism of these analyses is under investigation. Our findings suggest that targeting HIKESHI may be a promising therapeutic strategy against refractory pancreatic cancer. Citation Format: Bilguun Erkhem-Ochir, Takehiko Yokobori, Gendensuren Dorjkhorloo, Haruka Okami, Takaomi Seki, Norifumi Harimoto, Ryo Muranushi, Kouki Hoshino, Kei Hagiwara, Norihiro Ishii, Mariko Tsukagoshi, Kenichiro Araki, Hiroshi Saeki, Ken Shirabe. Nuclear transporter HIKESHI is a novel independent poor overall survival for pancreatic cancer; targeting can inhibit pancreatic cancer growth via HSP70 suppression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB274.

Senolysis of gemcitabine-induced senescent human pancreatic cancer cells.

Gemcitabine (GEM) is often used to treat pancreatic cancer. Many anti-cancer drugs induce cancer cell death, but some cells survive after cell cycle arrest. Such a response to DNA damage is termed cellular senescence. Certain drugs, including the Bcl-2-family inhibitor ABT-263, kill senescent cells; this is termed senolysis. In this study, we examined the therapeutic benefits of ABT-263 in GEM-induced senescence of human pancreatic cancer cells. Of four pancreatic cancer cell lines (PANC-1, AsPC-1, CFPAC-1, and PANC10.05), GEM induced senescent features in PANC-1 and AsPC-1 cells, including increases in the cell sizes and expression levels of mRNAs encoding interleukin (IL)-6/IL-8 and induction of β-galactosidase. Successive treatment with GEM and ABT-263 triggered apoptosis in PANC-1 and AsPC-1 cells and suppressed colony formation significantly. Senolysis of GEM-induced senescent pancreatic cancer cells by ABT-263 was triggered by a Bcl-xL inhibitor, but not by a Bcl-2 inhibitor, suggesting a central role for Bcl-xL in senolysis. In a xenograft mouse model, combined treatment with GEM and ABT-737 (an ABT-263 analog exhibiting the same specificity) suppressed in vivo growth of AsPC-1 significantly. Together, our results indicate that sequential treatment with GEM and senolytic drugs effectively kill human pancreatic cancer cells.

MSLN induced EMT, cancer stem cell traits and chemotherapy resistance of pancreatic cancer cells

Chemoresistance is one of the main reasons for poor prognosis of pancreatic cancer. The effects of mesothelin (MSLN) on chemoresistance in pancreatic cancer are still unclear. We aim to investigate potential roles of MSLN in chemoresistance and its relationship with proliferation, epithelial-mesenchymal transition (EMT) and cancer stemness of pancreatic cancer cells. Human pancreatic cancer cell lines ASPC-1 and Mia PaCa-2 with high and low expression of MSLN, respectively, were selected. The ASPC-1 with MSLN knockout (KO) and Mia PaCa-2 of MSLN overexpression (OE) were generated. The effects of MSLN on cell phenotypes, expression of EMT-related markers, clone formation, tumor sphere formation, and pathologic role of MSLN in tumorigenesis were detected. Sensitivity of tumor cells to gemcitabine was evaluated. The results showed that adhesion, proliferation, migration and invasion were decreased significantly in ASPC-1 with MSLN KO, whereas increased significantly in Mia PaCa-2 with MSLN OE. The size and the number of clones and tumor spheres were decreased in ASPC-1 with MSLN KO, and increased in Mia PaCa-2 with MSLN OE. In xenograft model, tumor volume was decreased (tumor grew slower) in MSLN KO group compared to control group, while increased in MSLN OE group. Mia PaCa-2 with MSLN OE had a higher IC50 of gemcitabine, while ASPC-1 with MSLN KO had a lower IC50. We concluded that MSLN could induce chemoresistance by enhancing migration, invasion, EMT and cancer stem cell traits of pancreatic cancer cells. Targeting MSLN could represent a promising therapeutic strategy for reversing EMT and chemoresistance in pancreatic cancer cells.

The DDX6/KIFC1 signaling axis, as regulated by YY1, contributes to the malignant behavior of pancreatic cancer.

Human DEAD/H box RNA helicase DDX6 acts as an oncogene in several different types of cancer, where it participates in RNA processing. Nevertheless, the role of DDX6 in pancreatic cancer (PC), together with the underlying mechanism, has yet to be fully elucidated. In the present study, compared with adjacent tissues, the level of DDX6 was abnormally increased in human PC tissues, and this increased level of expression was associated with poor prognosis. Furthermore, the role of DDX6 in PC was investigated by overexpressing or silencing the DDX6 in the PC cell lines, SW1990 and PaTu-8988t. A xenograft model was established by injecting nude mice with either DDX6-overexpressing or DDX6-silenced SW1990 cells. DDX6 overexpression promoted the proliferation and cell cycle transition, inhibited the cell apoptosis of PC cells, and accelerated tumor formation, whereas DDX6 knockdown elicited the opposite effects. DDX6 exerted positive effects on PC. RNA immunoprecipitation assay showed that DDX6 bound to kinesin family member C1 (KIFC1) mRNA, which was further confirmed by RNA pull-down assay. These results suggested that DDX6 positively regulated the expression of KIFC1. KIFC1 overexpression enhanced the proliferative capability of PC cells with DDX6 knockdown and inhibited their apoptosis. By contrast, DDX6 overexpression reversed the inhibitory effect of KIFC1 silencing on tumor proliferation. Subsequently, the transcription factor Yin Yang 1 (YY1) was shown to negatively regulate DDX6 at both the mRNA and protein levels. Dual-luciferase reporter assay verified that YY1 targeted the promoter of DDX6 and inhibited its transcription. High expression levels of YY1 decreased the proliferation of PC cells and promoted cell apoptosis, although these effects were reversed by DDX6 overexpression. Taken together, YY1 may target the DDX6/KIFC1 axis, thereby negatively regulating its expression, leading to an inhibitory effect on pancreatic tumor.

Synchrotron radiation-FTIR biomolecular assessment in PANC-1 and MIA PaCa-2 pancreatic cancer cells after treatment with Gd-TiO2 nanoparticles

The increasing incidence rate of pancreatic cancer worldwide, along with high resistance to existing chemotherapies and radiotherapy, demand new approaches for its treatment. Nanoparticle-guided cancer treatment modalities are promising in their anti-cancer efficiency and upgrading potential. We have synthesized and characterized Gd-doped titanium-dioxide nanoparticles (Gd-TiO2 NPs), which showed good biocompatibility with MIA PaCa-2 and PANC-1 pancreatic cancer cell lines. To assess the effect that these NPs have on tumor cells, we used synchrotron radiation Fourier transform infrared spectroscopy and principal component analysis. We elucidated the biochemical changes in the two cell lines induced after their interaction with these NPs. Results demonstrated changes in nucleic acids and carbohydrate metabolism and alterations in the protein conformation in both cell lines. In addition, considerable changes in lipid area were observed in PANC-1 cell line, suggesting that Gd-TiO2 NPs have a prospective to be further studied as systems for pancreatic cancer treatment.

Hyperthermia improves gemcitabine sensitivity of pancreatic cancer cells by suppressing the EFNA4/β-catenin axis and activating dCK

BackgroundPreviously, our investigations have underscored the potential of hyperthermia to improve the therapeutic efficacy of gemcitabine (GEM) in pancreatic cancer (PC). Nonetheless, the precise underlying mechanisms remain elusive. MethodsWe engineered two GEM-resistant PC cell lines (BxPC-3/GEM and PANC-1/GEM) and treated them with GEM alongside hyperthermia. The impact of hyperthermia on the therapeutic potency of GEM was ascertained through MTT assay, assessment of the concentration of its active metabolite dFdCTP, and evaluation of deoxycytidine kinase (dCK) activity. Lentivirus-mediated dCK silencing was further employed to validate its involvement in mediating the GEM-sensitizing effect of hyperthermia. The mechanism underlying hyperthermia-mediated dCK activation was explored using bioinformatics analyses. The interplay between hyperthermia and the ephrin A4 (EFNA4)/β-catenin/dCK axis was investigated, and their roles in GEM resistance was further explored via the establishment of xenograft tumor models in nude mice. ResultsHyperthermia restored dCK expression in GEM-resistant cell lines, concurrently enhancing GEM sensitivity and fostering DNA damage and cell death. These observed effects were negated by dCK silencing. Regarding the mechanism, hyperthermia activated dCK by downregulating EFNA4 expression and mitigating β-catenin activation. Overexpression of EFNA4 activated the β-catenin while suppressing dCK, thus diminishing cellular GEM sensitivity—a phenomenon remediated by the β-catenin antagonist MSAB. Consistently, in vivo, hyperthermia augmented the therapeutic efficacy of GEM on xenograft tumors through modulation of the ephrin A4/β-catenin/dCK axis. ConclusionThis study delineates the role of hyperthermia in enhancing GEM sensitivity of PC cells, primarily mediated through the suppression of the EFNA4/β-catenin axis and activation of dCK.

Extracts of Cistus creticus cultivated at different salinity levels exhibit promising therapeutic potential for pancreatic cancer cell lines

BackgroundCistus creticus L. subsp. creticus is a characteristic species commonly distributed in the Mediterranean region. It has been proved to contain antibacterial, antifungal, anti-inflammatory, and anti-cancer traits. PurposeIn this study, C. creticus plants were grown in hydroponic culture under different salinity levels (0, 30, or 60 mM NaCl) and were extracted using two other solvents (70 % ethanol and distilled water). The effect of salinity on plant growth and mineral homeostasis, biochemical content of extracts and therapeutic impact on cell death and cell survival in pancreatic cancer cell lines were studied. MethodsPancreatic cancer cell lines (PANC-1, MIA PaCa-2, and AsPC-1) were treated with different C. creticus extracts were grown, and cell survival, autophagy and apoptosis signaling pathways were measured. ResultsEspecially, ethanol extraction of C. creticus showed a remarkably therapeutic effect on cell death and cell survival. Ethanol extracts have higher polyphenolic content and are more effective than water extracts. It has been found that C. creticus is an important mitochondria-targeted antioxidant in pancreatic cancer cells. ConclusionThis research could be important to evaluate how stress physiology relates to the chemical composition and the therapeutic effectiveness of C. creticus.

Abstract 2431: Identification of highly expressed exosomal RNAs derived from pancreatic cancer cell lines and exploration of their potential as biomarkers

Abstract Background: Not all exosomes in the blood of cancer patients originate from cancer cells. This study evaluated the relationships between mRNAs and miRNAs in pancreatic cancer (PC) cell lines and those in exosomes secreted into the supernatant. We aimed to identify candidates for PC biomarkers from cancer-specific exosomal mRNAs and miRNAs. Materials and Methods: Three PC cell lines were utilized. Exosomes were isolated from the supernatant, and cellular and exosomal mRNAs and miRNAs were extracted. Subsequently, RNA sequencing (RNA-seq) was performed, and the data were analyzed using R software. Results: In the MIAPaCa-2 cell line, the expression of cellular mRNAs and corresponding exosomal mRNAs showed a strong correlation. In the other two cell lines, a moderate correlation was observed. Across all three cell lines, cellular miRNAs and corresponding exosomal miRNAs exhibited moderate correlation. We focused on RNAs that were lowly expressed in cells but highly expressed in exosomes and conducted differential expression analysis using R. Compared with healthy volunteers in the GEO database, some RNAs showed high expression in the supernatant and low expression in the plasma of healthy volunteers. Conclusion: We identified three mRNAs and three miRNAs that may serve as promising biomarker candidates for PC. Citation Format: Yuhan Rong, Naoki Okubo, Noritoshi Kobayashi, Eriko Katsuta, Yasushi Ichikawa. Identification of highly expressed exosomal RNAs derived from pancreatic cancer cell lines and exploration of their potential as biomarkers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2431.

Abstract 5685: MiR-345-5p attenuates pancreatic cancer metastasis through KLK7/E-Cad/β-Catenin/Rap1 axis

Abstract Background: Pancreatic cancer (PC) is a highly lethal malignancy with a 5-year survival rate of 10.8%, primarily attributed to early invasion and metastasis. Epithelial-to-mesenchymal transition (EMT) plays a crucial role in the metastatic progression of PC. Existing therapies, such as Gemcitabine, Abraxane, and FOLFIRINOX, have limited efficacy, emphasizing the need for identifying molecular targets to enhance PC patient survival. Recent advancements in miRNA therapies show promise, but a detailed understanding of miRNAs regulating PC growth and metastasis remains elusive. This study aims to uncover the role of miR-345-5p in regulating PC metastasis. Methods: A miRNA microarray analysis of KC mouse data identified miR-345-5P as one of the significantly down-regulated miRNA, targeting multiple metastasis-related genes. Expression of miR-345-5p in PC was assessed using published databases, validated through qPCR and ISH in PC cell lines, serum samples, and patient tissue. In vitro overexpression and in vivo assays elucidated the functional roles of miR-345-5p in PC metastasis. A computational approach and dual-luciferase assays were used to identify the target of miR-345-5p. Western blots, immunohistochemistry, and immunofluorescence were utilized to decipher miR-345-5p mediated molecular mechanisms. Results: MiR-345-5p expression was downregulated in PC tissue, serum, and cell lines compared to non-tumor tissues. Restoring miR-345-5p reduced PC cell proliferation, migration, and invasion in vitro and in vivo. KLK 7 was confirmed as a direct target, and functional experiments revealed miR-345-5p's acting as a tumor-suppressor by inhibiting the E-Cad/B-catenin/Rap1 axis signaling pathway. MiR-345-5p overexpression stabilized the E-Cadherin-β-catenin complex, leading to reduced β-catenin accumulation in the nucleus and thus leading to declined transcription of crucial genes (C-myc, cyclin D1 & MMP7) responsible for PC progression. Given the pivotal role of the c-myc gene in metabolism, we performed a metabolomic analysis. The findings unveiled significant metabolic rewiring, marked by reductions in glucose and amino acid metabolism, attributable to decreased C-myc expression due to restoration of miR-345 in PC cells. Conclusion: MiR-345-5p overexpression negatively regulates PC proliferation and EMT by directly targeting KLK7/E-Cad/β-catenin/Rap1 axis. Restoring miR-345-5p with standard care therapies could be a promising strategy for improving survival in metastatic PC patients. Citation Format: Nagabhishek Sirpu Natesh, Susheel Kumar Nethi, Brianna M White, Jussuf T. Kaifi, Surinder K. Batra, Surya Mallapragada, Satyanarayana Rachagani. MiR-345-5p attenuates pancreatic cancer metastasis through KLK7/E-Cad/β-Catenin/Rap1 axis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5685.

Abstract 5876: ALDH3A1 as a predictive marker for gemcitabine treatment in pancreatic cancer

Abstract Background: Pancreatic cancer is one of the incurable cancer. The currently preferred chemotherapy regimens, including gemcitabine, do not extend patient survival by more than a year, and studies on drug resistance mechanisms or biomarkers for the drug sensitivity are very limited. Aldehyde dehydrogenase (ALDH) is a family of enzymes that play an essential role in cell survival and protection. In cancer cells, it is thought that overexpression of the ALDHs confers a survival advantage to the cells, which is associated with a poor prognosis. In this study, we analyzed the drug sensitivity between ALDH3A1 expression and gemcitabine in PDAC cells. Method: In this study, we investigated the relationship between ALDH3A1 expression and gemcitabine sensitivity using 8 pancreatic cancer cell line. We observed changes in the pattern of ALDH3A1 expression in established gemcitabine resistance pancreatic cancer cell line. Following this, we functionally inhibited ALDH3A1 to determine its impact on gemcitabine sensitivity using shRNA method. To elucidate the molecular mechanisms related with ALDH3A1, we performed multi-omics analysis such as transcriptomics and proteomics. Results: As a results, there was a negative correlation between ALDH3A1 expression (Protein & mRNA) and gemcitabine resistance 8 pancreatic cancer cell lines. However, in established gemcitabine resistance BxPC3 cells compared to control cells, gemcitabine resistance increased more than 1000 fold while the protein expression was decreased. In these results correlated with inhibition of ALDH3A1 expression using shALDH3A1 experiments both change ALDH3A1 expression and gemcitabine resistance. In biological phenotype experiments, cell proliferation and migration increased in the ALDH3A1 inhibition groups. When using combination treatment of gemcitabine and CB29, a known ALDH3A1 inhibitor, drug resistance increased by more than 2,000 fold in the combination treatment groups. In RNAseq analysis, Adherens junction, Ras signaling, Axon guidance, and JAK-STAT signaling pathway related genes were up regulated in shALDH3A1 treatment group compared control. Conclusion: In this study, we demonstrated the negative correlation between ALDH3A1 and gemcitabine resistant in pancreatic cancer cell lines and suggesting that it may serves as a predictive marker for gemcitabine treatment in pancreatic cancer patients. Keywords: Pancreatic ductal adenocarcinoma (PDAC), Aldehyde dehydrogenase 3 family member A1 (ALDH3A1), Gemcitabine sensitivity. Citation Format: Jin Su Kim, Ji Young Keum, Chan Hee Park, Seungmin Bang. ALDH3A1 as a predictive marker for gemcitabine treatment in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5876.

Abstract 1794: PAF1 reprograms metabolism in pancreatic cancer by interacting with HIF1α

Abstract Background: Reprogrammed cellular metabolism has been refocused on in the past decade, as many cancer cells rewire various metabolic pathways to facilitate their survival, unlimited cell growth, and division. RNA Polymerase II-Associated Factor 1 (PAF1)/Pancreatic Differentiation 2 (PD2) is a core subunit of the human PAF1 Complex (PAF1C) that regulates the RNA polymerase II function during transcriptional elongation in normal cells; however, its overexpression has been implicated in promoting pancreatic tumorigenesis and metastasis through epigenetic regulation, chemoresistance, radioresistance and maintenance of cancer stem cells. While most of these studies have provided evidence of the multifunctional nature of PAF1/PD2 in PDAC progression, no study has investigated the role of PAF1/PD2 during the metabolic rewiring of cancer. Methods: In this study, we sought to examine the role of PAF1/PD2 in the metabolic rewiring of cancer cells in pancreatic ductal adenocarcinoma (PDAC) and decipher how PAF1/PD2 mediates this metabolic reprogramming in PDAC. Pancreatic cancer cell lines were transfected with shRNAs to knock down PAF1/PD2. Metabolic genes regulated by PAF1/PD2 were identified by qPCR and western blot upon PAF1/PD2 depletion. Metabolic assays were performed to investigate the role of PAF1/PD2. Immunoprecipitations identified proteins that interact with PAF1/PD2. Confocal microscopy confirmed the co-localization of PAF1/PD2 with its protein partners. We performed chromatin immunoprecipitation (ChIP) - Polymerase chain reaction to confirm the binding of the PAF1 sub-complex to its target gene. Results: Our results showed that pancreatic cancer cells depleted PAF1/PD2 downregulate genes involved in aerobic glycolysis compared to control cells. Also, the lactate release assay indicated that more lactate was produced in control cells than in cells with PAF1/PD2 knockdown. Interestingly, we identified that HIF1α interacts with PAF1, specifically in pancreatic cancer cells. PAF1 and HIF1α co-localization was observed in pancreatic cancer cell lines. We also observed that the PAF1/PD2- HIF1α complex bound to the promoter region of LDHA to regulate the expression of LDHA in pancreatic cancer cells, reprogramming the metabolism to utilize the aerobic glycolysis pathway preferentially. Conclusions: In conclusion, our study shows that PAF1/PD2 rewires the metabolism of PDAC by interacting with HIF1 α to regulate the expression of LDHA, which is the rate-limiting step of aerobic glycolysis. Citation Format: Ayoola O. Ogunleye, Neelanjana Gayen, Saravanakumar Marimuthu, Rama Krishna Nimmakayala, Sanchita Rauth, Zahraa Alsafwani, Jesse L. Cox, Surinder K. Batra, Moorthy P. Ponnusamy. PAF1 reprograms metabolism in pancreatic cancer by interacting with HIF1α [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1794.

Abstract 1828: Development and evaluation of novel gemcitabine analog for the treatment of pancreatic cancer

Abstract Purpose: Gemcitabine acts against pancreatic cancer and a wide range of solid tumors and it is known to be rapidly deaminated or metabolized in the blood to an inactive metabolite. To increase its therapeutic levels, gemcitabine is administered at high doses causing severe side effects. To improve its metabolic stability, increase cytotoxic activity, and limit the phenomena of resistance many alternatives have emerged, such as modifying gemcitabine. The purpose of this study was to develop and characterize a novel gemcitabine analog with improved stability and enhanced anticancer activity against pancreatic cancer. Methods: A novel analog of gemcitabine (GemAGY) was designed and synthesized through chemical modification of gemcitabine hydrochloride (GemHCl) at position 4 with hydroxylamine. Gem AGY was characterized using nuclear magnetic resonance (NMR), micro-elemental analysis, and purity determined using High-Performance Liquid Chromatography (HPLC). Pancreatic cancer cell lines (MiaPaCa-2, BxPC3, and PANC-1) were treated with GemHCl and GemAGY. Cytotoxicity, cell migration, clonogenic assay, cell cycle, and apoptosis studies were performed to evaluate the effect of GemAGY against pancreatic cancer cell lines. Results: The percent purity of GemAGY was over 99.6%. GemAGY demonstrated a higher cytotoxic effect against pancreatic cancer cells with a lower half-maximal inhibitory concentration (IC50) value (2.5-fold high) compared with GemHCl. For MiaPaCa-2 cells, the IC50 value of GemAGY was found to be 1.63 ± 0.15 µm compared to GemHCl (4.0 ± 1.7 µm). GemAGY also exhibited higher cytotoxicity in PANC-1 (IC50: 1.7 ± 0.2µm) compared to GemHCl (IC50: 5.6 ±1.3 µm) and the IC50 value of GemAGY (3.2±1.1µm) in BxPC-3 cells was significantly better than GemHCl (5.5±1.0 µm). The cell cycle analysis indicated that GemAGY-treated MiaPaCa-2 cells at 1.50 µm concentration had a higher G0/G1 population (60.24%) compared to GemHCl-treated cells (55%). The cell migration studies showed that GemAGY-treated MiaPaCa-2 cells at 12.5 µm concentration significantly inhibited cell mobility towards the wound area with migrated cells found to be 65 ± 6 compared to Gem HCl (174.3 ± 4.0) (GemAGY vs GemHCl; p<0.0001). Similarly, GemAGY-treated PANC-1 cells at 12.5 µm concentration significantly inhibited cell mobility towards the wound area with (98 ± 5.1) compared to Gem HCl (254 ± 6.6). The quantitative analysis of apoptosis studies showed that GemAGY significantly induced higher apoptosis in a dose-dependent manner in MiaPaCa-2 cells compared to GemHCl as concentration increases (GemAGY vs GemHCl; p <0.0001). Conclusion: This study demonstrates that GemAGY increases the anticancer activity of gemcitabine and may have the potential to improve the chemotherapeutic efficacy of gemcitabine in the treatment of pancreatic cancer. Citation Format: Joy Okoro, Raviteja Bulusu, Esther K. Frimpong, Sherise Rodgers, Bo Han, Xue Zhu, Edward Agyare. Development and evaluation of novel gemcitabine analog for the treatment of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1828.

Abstract 3141: LCB02A an antibody drug conjugate (ADC) targeting CLDN18.2 expressing solid tumors

Abstract Claudin18.2 (CLDN18.2) is an isoform of CLDN18 with very limited expression in gastric epithelial cells in normal tissues, and aberrant overexpression in gastric, pancreatic, esophageal, ovarian, lung, and other solid tumors. This expression pattern presents a significant hurdle in developing an ADC targeting CLDN18.2 due to potential on-target toxicity to the gastrointestinal (GI) tract. LCB02A is an antibody-drug conjugate (ADC) that targets CLDN18.2, composed of a topoisomerase 1 inhibitor (TOPO1i) as payload (DAR4) linked to the CLDN18.2 specific binding PR301839 hClaudin18.2 (CLDN18.2) is an isoform of CLDN18 with very limited expression in gastric epithelial cells in normal tissues, and aberrant overexpression in gastric, pancreatic, esophageal, ovarian, lung, and other solid tumors. This expression pattern presents a significant hurdle in developing an ADC targeting CLDN18.2 due to potential on-target toxicity to the gastrointestinal (GI) tract. LCB02A is an antibody-drug conjugate (ADC) that targets CLDN18.2, composed of a topoisomerase 1 inhibitor (TOPO1i) as payload (DAR4) linked to the CLDN18.2 specific binding PR301839 humanized IgG1 antibody utilizing LegoChem Biosciences’ ConjuAllTM technology. LCB02A showed potent in vitro cytotoxicity in CLDN18.2 expressing gastric and pancreatic cancer cell lines. LCB02A also showed excellent anti-tumor efficacy in various gastric and pancreatic cell line-derived xenograft (CDX) models and patient-derived xenograft (PDX) models. Preliminary toxicity assessment in cynomolgus monkeys (CLDN18.2 cross-reactive species) demonstrated that LCB02A was well tolerated, with repeat doses of up to 60 mg/kg. In conclusion, LCB02A is a promising ADC for the treatment of CLDN18.2 expressing solid tumors including gastric cancer (GC) and pancreatic ductal adenocarcinoma (PDAC). umanized IgG1 antibody utilizing LegoChem Biosciences’ ConjuAllTM technology. LCB02A showed potent in vitro cytotoxicity in CLDN18.2 expressing gastric and pancreatic cancer cell lines. LCB02A also showed excellent anti-tumor efficacy in various gastric and pancreatic cell line-derived xenograft (CDX) models and patient-derived xenograft (PDX) models. Preliminary toxicity assessment in cynomolgus monkeys (CLDN18.2 cross-reactive species) demonstrated that LCB02A was well tolerated, with repeat doses of up to 60 mg/kg. In conclusion, LCB02A is a promising ADC for the treatment of CLDN18.2 expressing solid tumors including gastric cancer (GC) and pancreatic ductal adenocarcinoma (PDAC). Citation Format: Eun ji Baek, Chul-Woong Chung, Changsik Park, Taeik Jang, Yeojin Jeong, Sang-Ah Lee. LCB02A an antibody drug conjugate (ADC) targeting CLDN18.2 expressing solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3141.

Abstract 6492: KRAS inhibitor-based combinations in pancreatic cancer identified through Optim.AI

Abstract PURPOSE: KRAS is the most frequently mutated oncogene in almost all cancers, including pancreatic ductal carcinoma (PDAC) and colorectal cancer. Increased development efforts targeting KRAS mutations have yielded recent FDA approvals of two KRAS G12C inhibitors. However, this is beneficial for only a small proportion of patients especially with indications like PDAC, where G12C mutations are rare (1%) while G12D mutations account for approximately 45%. Hence, there is a need to develop methods to target a broader range of KRAS mutations. Beyond direct gene mutation, activation of oncogenic KRAS and downstream or parallel signaling through other genomic alterations may benefit from KRAS-specific therapeutic strategies, especially combination therapy. In this study, Optim.AI™, a hybrid computational-experimental platform is harnessed to identify mutation-specific optimal therapies and to stratify efficacious drug combinations containing KRAS inhibitors in pancreatic cancer. METHODS: Optim.AI™ utilizes small datasets to rationally converge upon optimal drug combinations within a defined drug search space. By mapping experimental data points to a second-order quadratic function, Optim.AI™ predicts cell killing efficacies for all possible combinations, independent of the background of the models utilized. In this study, pancreatic cancer cell lines, with varying KRAS mutations, were tested with an array of 155 different combinations consisting of 12 drugs at varying concentrations. The drugs include KRAS G12C inhibitors, Sotorasib and Adagrasib, KRAS inhibitors in development, and approved therapies used clinically for PDAC. Post-drug treatment viability was measured and used for Optim.AI™ analysis, to rank and compare the top therapies across the different cell lines. RESULTS: Through Optim.AI™ analysis, we observed a range of sensitivities towards KRAS inhibition, even for both Sotorasib and Adagrasib. Compared against HT-29 cell line, harboring wild type KRAS, we have identified distinct combinations of KRAS inhibitors with standard of care drugs on cell lines with G12D, G12V and G12C KRAS mutants. Subsequent validation of the top and bottom ranked combinations for the respective cell lines correspond to the predicted Optim.AI™ results. CONCLUSIONS: This study has demonstrated the differential sensitivities of the KRAS inhibitors across several pancreatic cancer cell lines, with different KRAS mutational profiles. Notably, we have identified KRAS inhibitor-based combinations that are distinctly different from wild type KRAS cell line. These preliminary findings could be used to drive precision medicine by identifying suitable biomarkers to potentially stratify pancreatic cancer patients and improve treatment outcomes. Future work includes evaluating the clinical significance of these combinations by validating them on patient-derived organoids. Citation Format: Jhin Jieh Lim, Edward Kai-Hua Chow, Masturah Rashid. KRAS inhibitor-based combinations in pancreatic cancer identified through Optim.AI [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6492.

Abstract 4375: ALPPL2 is involved in MAPK signal pathway of pancreatic cancer

Abstract Background: Alkaline phosphatase placental-like 2 (ALPPL2) is a member of the alkaline phosphatase family. ALPPL2 is currently being investigated as a novel therapeutic target due to its cancer-specific expression. However, our knowledge of ALPPL2 biological function in cancer is limited. Here we describe an interaction between ALPPL2 and MAPK signaling pathway in pancreatic cancer. Materials and Methods: ALPPL2 mRNA expression in a panel of 27 pancreatic cell lines was determined by RNAseq. Protein expression and phosphorylation were measured by Western blot. ALPPL2 or ALPP were overexpressed in pancreas cell lines through a Lentiviral packaging system. MEK inhibitor (trametinib) activity was assessed in the same panel of cell lines using a 6-day cell proliferation assay on a Synentec Cellavista imaging system. MEK inhibitor resistant cell lines were developed by culturing cells in increasing concentrations of MEK162. Results: We hypothesized that pancreatic cancer cells with high ALPPL2 expression may be less sensitive to MEK inhibition. Our study from a panel of 27 pancreatic cancer cell lines showed that there was no correlation between ALPPL2 expression and response to trametinib. Western blot showed that the level of phospho-ERK was reduced in PANC1 and HPAC cells at 6 hours after trametinib treatment. Signaling rebounded at 24 and 48 hours, though it remained below baseline. Surprisingly, ALPPL2 expression increased at 24 and 48 hours after trametinib treatment, however, the mRNA of ALPPL2 and ALPP was down-regulated in three pancreatic cancer cell lines conditioned to be MEK inhibitor resistant. To understand the role of ALPPL2 in the MAPK signaling pathway in response to MEK inhibition, ALPPL2 and ALPP were overexpressed in 2 pancreatic cancer cell lines: YAPC and PSN-1 whose ALPPL2 and ALPP were undetected by Western blot prior to transfection. Overexpression of ALPPL2 or ALPP in YAPC cells resulted in more ERK phosphorylation with no change in total ERK, it also resulted in increased total and phospho-S6, and decreased phospho-STAT3 with no change in total STAT3. The amount of total and phospho-ERK were not changed in PSN-1 with overexpression of ALPPL2 or ALPP, however, it did result in increased total and phospho-S6, decreased phospho-STAT3. Interestingly, the expression of ALPPL2 and ALPP in YAPC cells with ALPPL2 or ALPP overexpression was increased at 48 hours after trametinib treatment. Discussion: To our knowledge, this is the first study to report that ALPPL2 interacts with the MAPK signaling pathway in pancreatic cancer cells. The activated MAPK signaling pathway appears to control ALPPL2 expression. The MAPK signaling pathway may regulate ALPPL2 expression through transcription and protein metabolism. We have also shown that ALPPL2 expression can activate MAPK signaling pathway. Our findings that ALPPL2 expression can be increased after trametinib treatment provide further rationale for targeting ALPPL2 in cancer. Citation Format: Ke Wei Gong, Bilal Hamid, Kenny W. Castro, Martina S. McDermott, Forrest Epstein, Kevin Chau, Chuhong Hu, Jun Zhang, Ming Lu, Benjamin G. Hoffstrom, Neil A. O'Brien, Dennis J. Slamon. ALPPL2 is involved in MAPK signal pathway of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4375.

Abstract 2093: Co-treatment with KRAS G12D inhibitor MRTX1133 plus TTFields against human pancreatic and Colorectal cancer cell lines results in synergistic up-regulation of cleaved PARP in KRAS G12D & unexpectedly in KRAS G12V as well

Abstract Background: Both colorectal and pancreatic cancers are major global health issues and among deadliest cancers with great disease burden. The most commonly mutated oncogenic alteration in human cancers is KRAS, which is prevalent in pancreatic malignancies. The KRAS G12D mutation subtype is present in more than 40% of pancreatic ductal adenocarcinoma (PDAC). To this date there is no available targeted therapy options for patients with KRAS subtype mutations. MRTX1133 has been identified as a non-covalent, potent, and selective inhibitor of KRASG12D. This small molecule inhibitor has been shown to suppress KRASG12D signaling in cells and in vivo. Tumor Treating Fields (TTFields) therapy is a novel approach to treating cancer, the electric fields alter the behavior of cancer cells and prevent them from growing and dividing. We hypothesize that the co-application of MRTX1133 with TTFields will enhance the effects of MRTX1133 in PDAC and colorectal cancer (CRC). Martials and Methods: PDAC cell line CFPAC1 with KRAS G12 V mutation and LS513 CRC cell line with KRAS G12D Mutation were treated with same dose of MRTX1133 (500 nM) and were co-treated with 150kHz TTFields. After 48 hours western blot was used to probe for cleaved PARP, cleaved C3, phosphor-ERK and DUSP6. Results: In both KRAS G12D & KRAS G12V cell lines, synergistic upregulation of cPARP was observed following 48-hour co-treatment. Synergistic inhibition of pERK happened in KRAS G12D cell line but no change in pERK level following TTFields or MRTX1133 treatment was seen. Upregulation of cC3 in KRAS G12D happened in only co-treatment conditions. DUSP6 levels increased following MRTX1133 treatment in KRAS G12V cell line. Conclusions: The surprising synergistic upregulation of cPARP in both KRAS G12V and G12D cells following co-treatment of KRAS G12D inhibitor MRTX1133 and TTFields can bring hope for other KRAS mutation subtypes as well as G12D. the increase in cPARP in KRAS G12V with no effect on pERK is particularly interesting and demonstrates the need of more studies to investigate the mechanisms of this synergy. Citation Format: Vida Tajiknia, Praveen Srinivasan, Maximilian PInho-Schwermann, William MacDonald, Connor Purcell, Wafik El-Deiry. Co-treatment with KRAS G12D inhibitor MRTX1133 plus TTFields against human pancreatic and Colorectal cancer cell lines results in synergistic up-regulation of cleaved PARP in KRAS G12D & unexpectedly in KRAS G12V as well [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2093.

Abstract 1645: Telomere-related genes associated with clinical-pathological in pancreatic ductal adenocarcinoma

Abstract Pancreatic ductal adenocarcinoma (PDAC) stands as one of the leading causes of cancer deaths worldwide. Studies have suggested that telomeres shortening is linked to genetic instability, higher risk of developing pancreatic cancer and pancreatic tumorigenesis. However, previous research has predominantly focused on either telomere length or specific telomere-related genes in pancreatic cancer. Therefore, it is essential to elucidate the association between telomere-related genes and pancreatic cancer to identify novel therapeutic strategies and establish prognostic biomarkers. Telomere-relevant genes were downloaded from the TelNet database. Expression data from primary tumour of The Cancer Genome Atlas-PAAD (TCGA-PAAD) cohort, the Genome Sequence Archive CRA001160 dataset and the GTEx database were obtained for screen for differentially expressed genes (DEGs). Gene set enrichment analisis was performed using the Enrichr software. The gene essentiality queried from the gene dependency database of the Achilles-DepMap project was also assessed. Finally, data of The Cancer Genome Atlas-PanCancer CaPa cohort and healthy pancreas from the GTEx database, obtained from the UCSC Xena project, were used to correlate gene expression with stemness signatures and clinical-pathological variables. We identified 300 telomere-associated DEGs, Of these, 73 genes were common to studies and they were to be enriched in biological processes related to the cell cycle and telomere homeostasis. The differentially expresed of ASS1, S100P, IGF2BP2, MET, ABCC3 and RAC1 genes were significantly associated with overall survival and recurrence-free survival, conferring them prognostic value. Furthermore, the expression profiles of these genes are significantly associated with the histological grade of the disease, as well as with tumour T-N classification, the presence of residual tumour, the degree of response to treatment and stemness signatures. Among these, three genes of particular interest (KLF3, RAC1 and ABI1) were identified and shown to be highly essential in pancreatic cancer cell lines. In summary, we have identified a telomere gene-related signature for pancreatic cancer, highlighting the prognostic value of KLF3, RAC1, and ABI1 genes. This signature is significantly associated with clinical-pathological variables of the disease, potentially offering new insights for individualized therapy. Citation Format: Erika Curiel Gómez, Vilma Maldonado Lagunas, Jorge Meléndez Zajgla. Telomere-related genes associated with clinical-pathological in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1645.