PDA Patients Research Articles

Speckle tracking of left ventricle in pediatric patients with hemodynamically significant patent ductus arteriosus (PDA): case–control study

BackgroundLeft ventricle (LV) volume overload is observed with hemodynamically significant PDA as it causes left to right shunting, leading to increased pulmonary blood flow. This overload causes LV remodeling as LV increases stroke volume in trial for compensation, but in larger shunts, patients may develop symptoms of congestive heart failure. Two-dimensional (2D) speckle-tracking echocardiography has emerged as a technique for objective and quantitative evaluation of global and regional myocardial function, independent of the angle of myocardial insonation.Aim of the workThis study aimed to evaluate left ventricular function by 2-dimensional speckle tracking in children with hemodynamically significant PDA.MethodsThis prospective controlled study was performed on 54 children divided into two groups (34 as cases and 20 as controls) to compare echocardiographic measurements. Conventional Echocardiography, tissue Doppler, and Speckle tracking were done for all patients, and measurements were compared.ResultsThere was a statistically significant difference (p value < 0.001) in global longitudinal strain (GLS) in the PDA group indicating a decrease in LV function in PDA patients. This difference was observed in several conventional echocardiographic parameters but not in tissue Doppler in our study.ConclusionLeft ventricle global strain is an important predictor of the myocardial performance index of the Left ventricle in patients with hemodynamically significant PDA and outweighs other conventional echocardiographic parameters and tissue Doppler indices.

Identification of potential biomarkers for pancreatic ductal adenocarcinoma: a bioinformatics analysis.

PDA is an aggressive cancer with a 5-year survival rate, which is very low. There is no effective prognosis or therapy for PDA because of the lack of target biomarkers. The objective of this article is to identify the target biomarkers for PDA using a bioinformatics approach. In this work, we have analysed the three microarray datasets from the NCBI GEO database. We used the Geo2R tool to analyse the microarray data with the Benjamini and Hochberg false discovery rate method, and the significance level cut-off was set to 0.05. We have identified 659 DEGs from the datasets. There are a total of 15 hub genes that were selected from the PPI network constructed using the STRING application. Furthermore, these 15 genes were evaluated on PDA patients using TCGA and GTEx databases in (GEPIA). The online tool DAVID was used to analyse the functional annotation information for the DEGs. The functional pathway enrichment was performed on the GO and KEGG. The hub genes were mainly enriched for cell division, chromosome segregation, protein binding and microtubule binding. Further, the gene alteration study was performed using the cBioportal tool and screened out six hub genes (ASPM, CENPF, BIRC5, TTK, DLGAP5, and TOP2A) with a high alteration rate in PDA samples. Furthermore, Kaplan-Meier survival analysis was performed on the six hub genes and identified poor-survival outcomes that may be involved in tumorigenesis and PDA development. So, this study concludes that, these six hub genes may be potential prognostic biomarkers for PDA.

Abstract 5553: Glycan CA19-9 mediated immune modulation in the pancreatic tumor microenvironment

Abstract Pancreatic ductal adenocarcinoma (PDA) is an intractable common malignancy with a pro-tumorigenic and immunosuppressive microenvironment (ME) that often limits treatment efficacy. Only 10-15% of patients are eligible for surgical resection, which is the only cure for PDA. The glycan CA19-9 is used to follow treatment response in PDA patients, but its functional role in PDA remained unknown until recently because rodents lack the ability to produce this carbohydrate. CA19-9 elevation in a KRAS-mutant background results in increased tumor proliferation and ME remodeling in mice. The tumor ME is encompassed by hypovascularized stroma consisting of cancer associated fibroblasts (CAFs) and tumor associated macrophages (TAMs) that contribute to desmoplasia and immunosuppression. All prior work investigating the tumor ME in PDA mouse models was performed in CA19-9 negative context, potentially missing a key element of PDA biology. Elevation of CA19-9 in mice caused expansion of both antigen-presenting (ap)CAFs and TAMs. CA19-9 induction in KRAS-mutant organoids increased gene expression of M-CSF, IL1a, and TGFb and led to increased apCAF and TAM differentiation. This project aims to uncover the mediators of CA19-9-mediated ME remodeling. CA19-9-modified macrophage colony-stimulating factor (M-CSF) was identified via immunoprecipitation and mass spectrometry from KRAS-mutant CA19-9-inducible pancreatic organoids. CA19-9-M-CSF induced increased immunosuppressive TAM differentiation in vivo and in vitro using a novel Macrophage, Organoid, and Fibroblast (MOrF) co-culture platform. This project will identify the mechanisms by which CA19-9 directly contributes to PDA ME immune modulation and unveil essential molecular underpinnings of PDA biology that will inform effective therapies in the future. Citation Format: Jasper Hsu, Tae Gyu Oh, Kristina L. Peck, Angelica E. Rock, Garret Bishop, Shira R. Okhovat, Susan M. Kaech, Dannielle D. Engle. Glycan CA19-9 mediated immune modulation in the pancreatic tumor microenvironment [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 5553.

Mucin 5 AC (MUC5AC) to predict pathologic treatment response (TR) to neoadjuvant chemotherapy (NAT) in resected-pancreatic ductal adenocarcinoma (R-PDA).

690 Background: MUC5AC is a gel-forming glycoprotein that exists in two major glycoforms in PDA. We studied the differential expression and predictive value of heavily glycosylated mature MUC5AC (MM) detected in the apical (Ap) and/or extracellular region (EC) and less-glycosylated immature MUC5AC (IM) detected only in the cytoplasm (Cy) in R-PDA. Methods: R-PDA formalin-fixed paraffin-embedded tissue blocks from January 2010 to June 2021 were obtained from the Ohio State University biorepository. Immunohistochemistry was performed to study the expression for MM and IM using monoclonal antibodies 45M1 and CLH2, respectively, and H-scores were calculated. Analysis of variance (ANOVA), t-tests, and Wilcoxon tests were used for statistical analysis. Results: Tissue samples from 100 R-PDA (43 received NAT and 57 had upfront surgery (UpS)) were available for testing. The median age of diagnosis was 65 years, and 50% were females. MM and IM were positive in 96% R-PDA (MM-95% Ap and 72% EC).In the NAT group, 89% (n=38) had 5-fluorouracil (5FU) based (36 FOLFIRINOX and 2 FOLFOX), and the rest received gemcitabine (Gem)-based therapy; patients received a median of 6 cycles of NAT; 37% (16) had preoperative chemoradiation (CRT) after NAT; 63% (27) had adjuvant systemic therapy (AST); pathologic objective response (OR = near complete or partial) and no response (NR) to NAT was reported in 63% (27) and 37% (16), respectively. In UpS group, 92% and 4% had AST (48 Gem-based, 4 5FU-based) and adjuvant CRT, respectively. The median time to recurrence (mTTR) and overall survival of the group was 15 and 33 months (m), respectively. NAT group had significantly (p&lt;0.05) lower EC-positive MM (mean H-score 108 vs. 151), IM (mean H-score 122 vs. 162), and mTTR (11 vs 15 m) than UpS group. IM levels were directly proportional to the response (near complete (mean H-score 98) &lt; partial (105) &lt; NR (152) &lt; UpS (162), p=0.04). Other TR results are discussed in the table. Conclusions: MUC5AC (IM &gt; MM) is a potential predictive marker for TR among PDA patients, specifically among those receiving FOLFIRINOX. These associations must be validated in further prospective studies.[Table: see text]

Nuclear GRP78 Promotes Metabolic Reprogramming and Therapeutic Resistance in Pancreatic Ductal Adenocarcinoma.

Stromal fibrosis limits nutritional supply and disarrays metabolism in pancreatic cancer (PDA, pancreatic ductal adenocarcinoma). Understanding of the molecular basis underlying metabolic cues would improve PDA management. The current study determined the interaction between glucose-regulated proteins 78 (GRP78) and hypoxia-inducible factor 1α (HIF-1α) and its mechanistic roles underlying PDA response to oxygen and glucose restrains. Gene expression and its association with clinicopathologic characteristics of patients with PDA and mouse models were analyzed using IHC. Protein expression and their regulation were measured by Western blot and immunoprecipitation analyses. Protein interactions were determined using gain- and loss-of-function assays and molecular methods, including chromatin immunoprecipitation, co-immunoprecipitation, and dual luciferase reporter. There was concomitant overexpression of both GRP78 and HIF-1α in human and mouse PDA tissues and cells. Glucose deprivation increased the expression of GRP78 and HIF-1α, particularly colocalization in nucleus. Induction of HIF-1α expression by glucose deprivation in PDA cells depended on the expression of and its own interaction with GRP78. Mechanistically, increased expression of both HIF-1α and LDHA under glucose deprivation was caused by the direct binding of GRP78 and HIF-1α protein complexes to the promoters of HIF-1α and LDHA genes and transactivation of their transcriptional activity. Protein complex of GRP78 and HIF-1α directly binds to HIF-1α own promoter and LDHA promoter, enhances the transcription of both HIF-1α and LDHA, whereas glucose deprivation increases GRP78 expression and further enhances HIF-1α and LDHA transcription. Therefore, crosstalk and integration of hypoxia- and hypoglycemia-responsive signaling critically impact PDA metabolic reprogramming and therapeutic resistance.

Risk stratification of hemodynamically significant patent ductus arteriosus by clinical and genetic factors.

Hemodynamically significant patent ductus arteriosus (hsPDA) is associated with increased comorbidities in neonates. Early evaluation of hsPDA risk is critical to implement individualized intervention. The aim of the study was to provide a powerful reference for the early identification of high-risk hsPDA population and early treatment decisions. We enrolled infants who were diagnosed with PDA and performed exome sequencing. The collapsing analyses were used to find the risk gene set (RGS) of hsPDA for model construction. The credibility of RGS was proven by RNA sequencing. Multivariate logistic regression was performed to establish models combining clinical and genetic features. The models were evaluated by area under the receiver operating curve (AUC) and decision curve analysis (DCA). In this retrospective cohort study of 2199 PDA patients, 549 (25.0%) infants were diagnosed with hsPDA. The model [all clinical characteristics selected by least absolute shrinkage and selection operator regression (all CCs)] based on six clinical variables was acquired within three days of life, including gestational age (GA), respiratory distress syndrome (RDS), the lowest platelet count, invasive mechanical ventilation, and positive inotropic and vasoactive drugs. It has an AUC of 0.790 [95% confidence interval (CI) = 0.749-0.832], while the simplified model (basic clinical characteristic model) including GA and RDS has an AUC of 0.753 (95% CI = 0.706-0.799). There was a certain consistency between RGS and differentially expressed genes of the ductus arteriosus in mice. The AUC of the models was improved by RGS, and the improvement was significant (all CCs vs. all CCs + RGS: 0.790 vs. 0.817, P < 0.001). DCA demonstrated that all models were clinically useful. Models based on clinical factors were developed to accurately stratify the risk of hsPDA in the first three days of life. Genetic features might further improve the model performance. Video Abstract (MP4 86834kb).

Functional and transcriptome analysis of peripheral blood lymphocytes in pancreatic cancer patients before and after chemotherapy.

Abstract Pancreatic Ductal Adenocarcinoma (PDA) is one of the most aggressive malignancies with a 5-year survival rate of 11% due to its high resistance to chemotherapy (CT). Understanding how to better induce an effector response following tumor associated antigen (TAA) stimulation is critical to set up an efficient tailored immunotherapy. Peripheral blood mononuclear cells (PBMC) from PDA patients, before and after gemcitabine-based CT, were in vitrostimulated with four TAA (ENO1, FUBP1, K2C8 and G3P) in presence or absence of immune-check point blockade (ICB). Gemcitabine increased the number of TAA recognized by patient T cells, which positively correlated with survival, a high-rate of TAA-induced proliferative responses and high IFN-γ production. By contrast, ICB suppressed TAA-induced proliferative responses before or after CT. Transcriptional analysis revealed that after CT only, responding patients displayed more differentially expressed genes than non-responding patients, suggesting that Gemcitabine modulates gene expression. The clonotypic analysis of ENO1-stimulated T cells from patients showed that TCRβ repertoire was modulated by CT as some clonotypes. These data indicate that Gemcitabine alone, as opposed to ICB, shifted the immunological tone toward a more effector phenotype. The expansion of V-J rearrangements due to TAA stimulation was enhanced by CT, with a stronger reactivity of precise TAA-specific T cell clones after CT, which support the development of precision immunotherapy in selected patient groups, based on CT combined with TAA vaccination. Supported by grants from AIRC (ID 19931)

Electrophysiological Signatures of Anxiety in Parkinson’s Disease (S51.008)

<h3>Objective:</h3> To assess the pattern of disturbances in the functional brain networks of Parkinson’s disease patients with anxiety (PD-A) compared to those without anxiety (PD-NA) and healthy controls (HC). <h3>Background:</h3> Anxiety is a common non motor symptom in PD occurring in up to 60% of the patients and affecting their quality of life. Despite their high prevalence, anxiety symptoms are often undiagnosed and untreated in PD. To date, functional and structural neuroimaging studies have contributed to our understanding of the motor and cognitive symptomatology of PD. Yet, the underlying pathophysiology of the anxiety symptoms in PD remains largely unknown and studies on their neural correlates are missing. <h3>Design/Methods:</h3> The present study comprised 68 non-demented PD patients and 25 healthy controls. PD patients were divided into two subgroups based on their Beck Anxiety Inventory (BAI) scale (cut-off=13): PD patients with anxiety (N=18) and without anxiety (N=50). Using their resting state high density electroencephalography (EEG) recordings at baseline, we assessed the frequency-dependent functional connectivity patterns that characterize the PD-A patients compared to both PD-NA patients and HC and we validated their relevance after 3 years. <h3>Results:</h3> We revealed different frequency-dependent pattern of connectivity that can characterize the PD-A group compared to PD-NA and HC: patterns with high functional connectivity were observed in delta, theta and gamma bands involving mainly fronto-temporal connections, whereas fronto-parietal patterns with low functional connectivity were pertinent in alpha and beta bands. Their corresponding network signature metric were strongly correlated with the anxiety scale of all the participants at baseline and after 3 years showing a predictive power of the revealed networks. <h3>Conclusions:</h3> Our findings suggest that PD-related anxiety can be characterized by a signature of different frequency-dependent pattern of disturbances in the functional brain networks. Resting state EEG could be a very promising biomarker for early detection of anxiety in PD. <b>Disclosure:</b> Mrs. Yassine has nothing to disclose. Mrs. Almarouk has nothing to disclose. Ute Gschwandtner has nothing to disclose. Dr. Auffret has received personal compensation for serving as an employee of France Développement Electronique. The institution of Dr. Auffret has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Britannia. Dr. Auffret has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Britannia. Dr. Auffret has received personal compensation in the range of $5,000-$9,999 for serving on a Speakers Bureau for Britannia. Dr. Auffret has received personal compensation in the range of $500-$4,999 for serving on a Speakers Bureau for Aguettant. The institution of Dr. Auffret has received research support from Association France Parkinson. The institution of Dr. Auffret has received research support from Homeperf. The institution of Dr. Auffret has received research support from LVL. The institution of Dr. Auffret has received research support from University of Rennes 1. The institution of Dr. Auffret has received research support from Aguettant. The institution of Dr. Auffret has received research support from Linde. The institution of Dr. Auffret has received research support from Plateforme Nationale pour la recherche sur la fin de vie. Dr. Auffret has received publishing royalties from a publication relating to health care. Mr. HASSAN has nothing to disclose. The institution of Dr. Fuhr has received research support from Roche. Mr. Verin has nothing to disclose.

Hemodynamic and Echocardiographic Characteristics and the Presence of Pulmonary Hypertension in Patent Ductus Arteriosus Patients who Underwent Transcatheter Closure.

We investigated the hemodynamic parameters of pediatric PDA patients and focused on the influence of PDA size on pulmonary arterial pressure and the prevalence of pulmonary hypertension. A total of 52 patients aged between 2months and 20years who received transcatheter closure of a PDA from January 2018 to June 2022 in our institution were retrospectively recruited. Their hemodynamic parameters collected both by echocardiography and by cardiac catheterization were analyzed to delineate the influence of PDA size on the pulmonary vascular system. The echocardiographic-based ductal size and indexed PDA size were 1.93mm (1.15-6mm) and 4.05mm/m2 (2.03-25.47mm/m2), respectively. The pulmonary artery pressure measured was 20.83mmHg (8-45mmHg). We found a positive correlation between indexed PDA size and mean pulmonary arterial pressure (mPAP) (Pearson correlation coefficient = 0.47, p < 0.001). A subgroup analysis showed that 28 patients (53.8%) developed pulmonary hypertension (PH) (defined as mPAP > 20mmHg). The median age of the PH group was 1.02years [range: 0.19-8.64], which was significantly younger than the non-PH group's median age of 3.43years [range: 0.42-19.96] (p = 0.001). The indexed PDA size for the PH group, 4.69mm/m2, was significantly higher than that of the non-PH group, 3.2mm/m2 (p = 0.004). The major risk factor for patients with PH was the PDA/BSA index, with an OR of 2.181 (95% CI, 1.224-3.887). Our demographic data showed younger patients with a higher PDA/BSA index are more likely to develop pulmonary hypertension.

Altered Fractional Amplitude of Low-Frequency Fluctuation in Anxious Parkinson's Disease.

Anxiety symptoms are persistent in Parkinson's disease (PD), but the underlying neural substrates are still unclear. In the current study, we aimed to explore the underlying neural mechanisms in PD patients with anxiety symptoms. 42 PD-A patients, 41 PD patients without anxiety symptoms (PD-NA), and 40 healthy controls (HCs) were recruited in the present study. All the subjects performed 3.0T fMRI scans. The fractional amplitude of low-frequency fluctuation (fALFF) analysis was used to investigate the alterations in neural activity among the three groups. A Pearson correlation analysis was performed between the altered fALFF value of the PD-A group and anxiety scores. Compared with HCs, PD-A patients had higher fALFF values in the left cerebellum, cerebellum posterior lobe, bilateral temporal cortex, and brainstem and lower fALFF values in the bilateral inferior gyrus, bilateral basal ganglia areas, and left inferior parietal lobule. Moreover, between the two PD groups, PD-A patients showed higher fALFF values in the right precuneus and lower fALFF values in the bilateral inferior gyrus, bilateral basal ganglia areas, left inferior parietal lobule, and left occipital lobe. Furthermore, Pearson's correlation analysis demonstrated that the right precuneus and left caudate were correlated with the Hamilton Anxiety Rating Scale scores. Our study found that anxiety symptoms in PD patients may be related to alterations of neurological activities in multiple brain regions. Furthermore, these may be critical radiological biomarkers for PD-A patients. Therefore, these findings can improve our understanding of the pathophysiological mechanisms underlying PD-A.

Regional activity alterations in Parkinson's disease patients with anxiety disorders: A resting-state functional magnetic resonance imaging study.

Previous studies have revealed alteration of functional connectivity (FC) in Parkinson's disease patients with anxiety (PD-A), but local brain activities associated with anxiety in Parkinson's disease (PD) patients remain to be elucidated. Regional homogeneity (ReHo) analysis was employed to investigate alterations of regional brain activities in PD-A patients. Resting-state functional magnetic resonance imaging (rs-fMRI) data were acquired from 42 PD-A patients, 41 PD patients without anxiety (PD-NA), and 40 age-and gender-matched healthy control (HC) subjects. ReHo analysis was used to investigate the synchronization of neuronal activities in brain regions in the three groups. The relationship between ReHo value and anxiety score in the PD-A group was also investigated. Parkinson's disease patients with anxiety showed increased ReHo values in the bilateral frontal lobes, caudate nucleus, and anterior cingulate gyrus [Gaussian random field (GRF) correction, voxel size p < 0.01, cluster size p < 0.05], compared with PD-NA patients and HC subjects, but the ReHo values of the right cerebellar hemisphere and posterior cerebellar lobe decreased (GRF correction, voxel size p < 0.01, cluster size p < 0.05). The increased ReHo values of the right superior frontal gyrus (r = 0.633, p = 0.001) and anterior cingulate gyrus (r = 0.45, p = 0.01) were positively correlated with anxiety scores in PD-A patients. The development of PD-A may be associated with dysfunctional local activities in multiple brain regions, including the frontal cortex, cerebella, basal ganglia, and limbic system. Abnormal ReHo values in these brain regions may serve as neuroimaging markers for the early diagnosis of PD-A. The results suggest that using ReHo analysis to identify functional changes in core regions may advance our understanding of the pathophysiological mechanisms underlying PD-A.

Abstract A036: Plasma extracellular RNA for the detection of pancreatic cancer

Abstract Pancreatic ductal adenocarcinoma (PDA) is expected to become the second leading cause of cancer-related death by 2030. Patients are often diagnosed at the metastatic stage, at which point treatments are limited and have non-uniform efficacy in patients. Advances in transcriptomic subtyping of tumor tissue can segregate patients into two broad subtypes, Classical and Basal-like, which are predictive of survival and response to first-line chemotherapy. However, tumor tissue is often difficult or impossible to obtain, particularly in patients with advanced disease, hindering the clinical implementation of transcriptomic subtyping. Profiling the extracellular RNA (exRNA) content in plasma provides an opportunity for a non-invasive readout of the tumor transcriptome. Here, we aim to establish the feasibility of liquid biopsy—particularly, plasma exRNA-profiling—for the diagnosis and subtyping of PDA. Extracellular vesicle (EV) encapsulated exRNA was isolated from 1mL of frozen plasma collected between 2017-2020 from a pilot cohort of 8 patients with stage III-IV PDA and processed alongside plasma from 6 healthy volunteers. UMI-tagged libraries were constructed from low-input exRNA to generate sequencing data. Gene body coverage analysis indicated that exRNAs consisted of intact full-length transcripts. Of these transcripts, we found that there was a median of 34,975 genes encoded by exRNAs, deriving mostly from protein-coding regions. Most genes captured by exRNA sequencing had low expression. Comparing between groups, we detected a significantly greater number of genes and novel splicing junctions in PDA patients. Of the cancer-specific transcripts, differential expression analysis revealed significant upregulation of membrane organelle and RNA-processing related pathways. PDA exRNA profiles did not completely recapitulate the bulk tumor transcriptome, suggesting specific mechanisms for EV encapsulation of RNAs and exRNA release. To classify tumor subtypes by plasma exRNA sequencing, we identified an exRNA-specific gene panel from bulk tumor transcriptome signatures. Using this gene panel, tumor subtype was correctly inferred from 6/7 PDA patients. A combined gene panel derived from all tumor-associated exRNA signatures distinguished 7/8 PDA patients from healthy controls. In conclusion, plasma exRNA recapitulated elements of the tumor transcriptome and has utility in PDA diagnosis. The establishment of diagnostic markers will be further investigated in the expanded study cohort comprised of 91 unique stage III-IV PDA donors with plasma collected at baseline, 150 stage III-IV PDA donors with plasma banked at timepoints throughout chemotherapy treatment, and 100 age- and sex-matched non-PDA controls donors. Leveraging the study cohort, future work will also investigate plasma exRNA signatures as prognostic markers to predict treatment outcomes and monitor treatment responses in patients with advanced PDA. Citation Format: Linsey Gong, Ling Xi, Yuanchang Fang, Karen Ng, Gun Ho Jang, Amy X. Zhang, Julie M. Wilson, Stephanie Ramotar, Jennifer J. Knox, Grainne M. O'Kane, Steven Gallinger, Hansen H. He, Faiyaz Notta. Plasma extracellular RNA for the detection of pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr A036.

Abstract C045: CA19-9-mediated remodeling of the pancreatic tumor microenvironment

Abstract Pancreatic ductal adenocarcinoma (PDA) is an intractable common malignancy with a pro-tumorigenic and immunosuppressive microenvironment (ME) that often limits treatment efficacy. The glycan CA19-9 is used to follow treatment response in PDA patients, but its functional role in PDA remained unknown until recently because rodents lack this carbohydrate. CA19-9 elevation in a KRAS-mutant background promotes rapid progression to invasive carcinoma and induces ME remodeling in mice. The tumor ME includes cancer associated fibroblasts (CAFs) and tumor associated macrophages (TAMs) that contribute to desmoplasia, immunosuppression, and therapeutic resistance. All prior work investigating the tumor ME in PDA mouse models was performed in a CA19-9 negative context, potentially missing a key element of PDA biology. Elevation of CA19-9 in mice caused expansion of both CAFs and TAMs, but the mechanisms by which CA19-9 contributes to ME remodeling remains largely unknown. We aim to uncover the direct and indirect mechanisms of CA19-9-mediated ME remodeling. We identified CA19-9 modified ligands by immunoprecipitation and mass spectrometry that may contribute to reprogramming of the PDA ME. In addition to exploring changes in abundance, localization, and protein interactions, we will functionally characterize these candidate effectors via CRISPR ablation and antibody blockade using a novel Macrophage, Organoid, and Fibroblast (MOrF) co-culture platform. MOrF co-cultures can be sustained for at least 10 days in defined media conditions and enable compartment specific delineation of paracrine and juxtacrine signals. While CA19-9 modification may play a direct role in stromal remodeling, we also investigate effectors that are altered in response to CA19-9 elevation. CA19-9 induction in KRAS-mutant organoid mono-cultures increased gene expression of IL1α, CSF1,and TGFβ. In addition, CA19-9 expression increased gene set enrichment of inflammatory programs inorganoid mono-culture and led to increased expression of CAF and TAM differentiation programs in MOrF co-culture. These results are being validated as discussed above as well as by evaluation in vivo in CA19-9 positive and negative syngeneic orthotopic transplantation and autochthonous genetically engineered mouse models using scRNA-seq, pharmacologic and genetic perturbation, as well as other approaches. Using these orthogonal approaches, we will identify the mechanisms by which CA19-9 directly and indirectly contributes to PDA ME remodeling and ultimately, contributes to treatment response in the future. Citation Format: Jasper Hsu, Sejin Chung, Angelica E. Rock, Shira R. Okhovat, Dannielle D. Engle. CA19-9-mediated remodeling of the pancreatic tumor microenvironment [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr C045.

Abstract A061: Utility of patient-derived organoids for precision pharmacotyping, phenotypic screens, and modeling of pancreatic disease

Abstract Pancreatic ductal adenocarcinoma (PDA) is projected to soon become the second leading cause of cancer death. Aberrant glycosylation is a hallmark of PDA, but the impact of glycosylation remains vastly understudied in PDA. Additionally, a lack of disease models that recapitulate glycosylation differences observed in human disease has hampered efforts to leverage glycan changes as therapeutic strategies. To address these needs, human patient-derived organoids (PDOs) were employed to address three major questions. First, the use of PDOs in drug screening experiments to characterize therapeutic sensitivity and resistance (i.e. “pharmacotyping”) is a promising precision medicine approach to inform clinical decisions. However, the extent to which different sources of basement membrane extracts (BMEs) in which PDOs are cultured contribute to organoid chemosensitivity profiles is unknown. Mouse and human PDA organoids representing a range of PDA disease stages (early, late, and metastasis) were subjected to a chemotherapy and targeted therapy pharmacotyping panel in different commercial BMEs. Differences in growth kinetics and drug response were analyzed. RNA-seq was performed to determine the impact of BME source on gene expression programs. Results from this work will provide key insights regarding optimal growth of pancreatic PDOs as well as the impact of exogenous factors on pharmacotyping results. Second, elevated serum levels of the glycan CA19-9 are observed in more than 75% of PDA patients. Recent evidence demonstrated that CA19-9 promotes pancreatitis and accelerates pancreatic tumorigenesis in mice, phenotypes that are reversible with the use of CA19-9-targeting antibodies. However, how CA19-9 synthesis is regulated is unknown. We developed organoid high-throughput screens to identify compounds that impact CA19-9 production. Continuing this screen across a library of hPDOs will lay the foundation for the identification of new therapeutic candidates and the understanding of regulators of glycosylation in PDA. Lastly, the pancreatitis is a major risk factor for the development of PDA, with hereditary pancreatitis patients having a 40-55% cumulative risk of developing PDA. However, reliable organoid models of human pancreatitis are lacking. To address this need, we optimized methods to derive PDOs from chronic pancreatitis specimens of genetic and non-genetic etiology. Human chronic pancreatitis organoids underwent multi-omic characterization and credentialing in order to provide the field with more representative models of pancreatitis in humans and a platform to explore early detection strategies and study pancreatic disease progression. Overall, we demonstrate the broad utility of PDOs to identify early detection biomarkers and targetable vulnerabilities in PDA. These models are a renewable resource for the field, enabling elucidation of the molecular underpinnings of the increased susceptibility of chronic pancreatitis to PDA and other fundamental aspects of PDA biology. Citation Format: Jan Lumibao, Shira Okhovat, Kristina Peck, Dannielle Engle. Utility of patient-derived organoids for precision pharmacotyping, phenotypic screens, and modeling of pancreatic disease [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr A061.

Effect of Intra Cardiac Shunts on Left Ventricular Systolic Function: A Speckle Tracking Study

Background: The left ventricular (LV) chamber size and its systolic function is the most common and quickest assessment made by echocardiography, either in the intra operative or intensive care setting, being the pressure generator for the blood supply to the body .Congenital cardiac defects come in two main types: atrial and ventricular septal defects (ASD/VSD). Regression or spontaneous closure may be the natural course of minor septal defects.&#x0D; Aim and Objectives: The aim of the study was to assess the feasibility of speckle tracking echocardiography in estimation of left ventricular systolic function in congenital shunt lesions (ASD, VSD and PDA).&#x0D; Subjects and Methods: This study was done in Tanta University Hospital including 270 patients. The patients were divided into four groups: ASD patients, VSD patients, PDA patients and control subjects.&#x0D; Results: It showed statistically significant difference between ASD, PDA and control group. The difference between the PDA group and the control group was statistically significant. There was a statistically significant difference between the ASD group and the control group in terms of EF percent, FS percent, and ESV ROC curve for Validity of GLS to predict LV systolic dysfunction in PDA Group. Sensitivity was 68 and sensitivity was 80. &#x0D; Conclusion: It was determined that Speckle-tracking echocardiography offers an additional non-invasive method for evaluating patients' left ventricular function. With congenital shunt lesions.

Abstract 1356: In vivo epigenetic CRISPR screen identifies PRMT5 as an immune suppressor in pancreatic cancer

Abstract Pancreatic ductal adenocarcinoma (PDA) is a devastating disease in which the mortality rate approaches the incidence rate. Desmoplasia is a hallmark of PDA, which creates a mechanical barrier surrounding the tumor cells, prevents appropriate vascularization and leads to pool immune infiltration. PDA exhibits a “cold” tumor microenvironment characterized by a prominent myeloid cell infiltration, which is extremely immune suppressive. Therefore, PDA has been demonstrated to be resistant to immunotherapy. Indeed, single-agent immune checkpoint inhibitor therapy has failed in clinical trials for PDA patients. Targeting epigenetic modification is an attractive approach to enable the response to immunotherapy in multiple solid tumor models. To identify epigenetic modulators as potential novel immunotherapeutic targets in PDA, we performed an orthotopic in vivo epigenetic CRISPR screen. Prmt5 was screened out as one of the top candidates. Further validation studies showed that Prmt5 knockdown (KD) alone or in combination with anti PD-1 treatment significantly inhibited the tumor progression. PRMT5 shRNA in combination with PD1 blockade can dramatically altered the tumor immune microenvironment (TME) with influx of effector CD8 T cells and changes in the macrophage/myeloid populations. Collectively, our work describes PRMT5 as an attracting target which governs tumor immunity in PDA. Citation Format: Hai Hu, Jiehui Deng, Fei Li, wei wang, Alireza Khodadadi-Jamayran. In vivo epigenetic CRISPR screen identifies PRMT5 as an immune suppressor in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1356.

Neoadjuvant and adjuvant antitumor vaccination alone or combination with PD1 blockade and CD137 agonism in patients with resectable pancreatic adenocarcinoma.

558 Background: Utilizing a vaccine that induces and activates host effector T cells and co-administering it with immune modulating agents that enhance anti-tumor T cell activity is a potential strategy for overcoming pancreatic adenocarcinoma’s (PDA) resistance to immunotherapy. Our prior clinical trial demonstrated a GM-CSF-secreting, allogeneic tumor cell vaccine (GVAX) increases infiltrating CD8+ T cells in PDA. Follow up preclinical work demonstrated therapeutic synergy between GVAX and PD-1inhibition (PD1) with efficacy further enhanced by CD137 agonism (CD137). Methods: This was a 3-arm trial of neoadjuvant &amp; adjuvant GVAX-based therapy in resectable (r) PDA patients (pts). Adults with clinically resectable, untreated PDA were enrolled in 1 of 3 study treatments: Arm A (GVAX alone), Arm B (GVAX + PD1 [Nivolumab]), or arm C (GVAX + PD1 + CD137 [Urelumab]). Treatment was given as follows: Day 1 - Cyclophosphamide 200mg/m2 IV (All Arms), Nivolumab 480mg IV (Arms B, C), Urelumab 8mg IV (Arm C); Day 2 – GVAX ID (All Arms). Pts were treated at 3 timepoints: 1) once 2 weeks prior to surgery; 2) once post-surgical recovery prior to standard of care adjuvant chemotherapy (SOC); 3) every month (up to 4 mo) following completion of SOC (if disease-free). SOC regimes included (m)FOLFIRINOX, Gem +/- Cap/NAB-Paclitaxel. The study was powered for a primary biologic endpoint: treatment-related change in intratumoral CD8+CD137+ T cells. Clinical endpoints included disease-free survival (DFS: time from surgery to recurrence), overall survival (OS: time from surgery to death), and safety. Results: 38 pts (N = 15 [Arm A], N = 13 [Arm B], N = 10 [Arm C]) were eligible for efficacy analysis (had R0/R1 resection) and 45 pts (N = 17 [A], N = 17 [B], N = 11 [C]) were eligible for safety analysis (had ≥1 dose of study treatment). Demographics, surgical pathology features, and SOC durations were similar in all Arms. At median follow up of 23 mo [A], 26 mo [B], and 22 mo [C], median DFS (95% CI) was 14.82 mo (6.0, NA), 16.23 mo (7.49, NA) and not reached (16.33, NA) for Arms A, B, C, respectively. There was no DFS benefit to adding PD1 compared to GVAX alone (HR 0.98 [95% CI 0.42, 2.27], p = 0.96). Combination CD137 + PD1 + GVAX was associated with marginally significant improved DFS compared to GVAX alone (HR 0.38 [95%CI 0.12, 1.19], p = 0.097) and GVAX + PD1 (HR 0.38 [95%CI 0.12, 1.21], p = 0.103). Median OS (95% CI) was 25.0 mo (18.8, NA), 26.4 mo (20.3, NA), and not yet reached for Arms A, B, C, respectively. There were no serious adverse events. In Arm C, 1 pt had grade 3 rash that delayed treatment and there was 1 instance of grade 2 AST/ALT elevation. The biologic endpoint will be reported at the meeting. Conclusions: Despite a small sample size, combining GVAX with dual immune-targeting of PD-1 blockade and CD137 agonism was safe and may enhance DFS in rPDA pts treated in the perioperative and post-adjuvant settings. Clinical trial information: NCT02451982.

Surgical Treatment for Patent Ductus Arteriosus: Our Experience of 12 Years.

IntroductionPatent ductus arteriosus (PDA) is a congenital heart disease that, if left untreated, can lead to pulmonary hypertension, congestive heart failure, and death. Here, we aimed to assess postoperative cardiac hemodynamic changes and surgical techniques, as well as early and late postoperative findings in surgically treated PDA patients.Materials and methodsWe retrospectively analyzed the data belonging to 126 patients whose PDA was surgically closed in our clinic from January 2001 to December 2012. With echocardiography being a standard in diagnosis and follow-up, angiography and computed tomography were also used in the presence of pulmonary hypertension and congenital heart disease, when needed. Postoperative data were compared between isolated PDA patients and those with congenital cardiac deformities.ResultsEvaluating the patients' pulmonary artery pressure (PAP), pulmonary hypertension was detected in 121 patients (96.0%). Preoperative PAP was significantly higher in PDA patients with congenital heart disease compared to the isolated PDA group (p<0.05). PAP decreased significantly in postoperative follow-up in both groups (p<0.05). However, this decrease was faster in the isolated PDA group than in patients with congenital heart disease and right-left shunt accompanying PDA (p<0.05). Regarding the correlation between ductus diameters and preoperative PAP, we found that as ductus diameter increased, PAP increased significantly (p<0.05).ConclusionsIn PDA patients, closing the ductus is necessary to prevent pulmonary and cardiac complications. Surgical closure remains one of the most effective methods for this, although there is little difference between surgical treatment methods in terms of mortality.

Metabolomics profiling reveals altered lipid metabolism and identifies a panel of lipid metabolites as biomarkers for Parkinson’s disease related anxiety disorder

ObjectivesAnxiety disorder is a common non-motor symptom in patient with Parkinson’s disease (PD). We aimed to explore its pathogenesis and identify plasma biomarkers using untargeted metabolomics analysis. MethodsConsecutive PD patients and healthy controls were recruited. Clinical data were assessed and patients with Parkinson’s disease related anxiety disorder (PDA) were recognized. Fast plasma samples were obtained and untargeted liquid chromatography-mass spectrometry-based metabolomics analysis was performed. Based on the differentially expressed metabolites from the above metabolomics analysis, correlation analyses and receiver operating characteristic curves (ROC) were further employed. ResultsAccording to the clinical data, PDA patients had lower plasma levels of total cholesterol, triglyceride, low-density lipoprotein cholesterol, and apolipoprotein B. There were thirty-nine differentially expressed metabolites in PDA patients when compared with the other two groups from the metabolomics analysis, respectively. Fourteen lipid metabolites were simultaneously altered between these two groups, and all of them were significantly decreased. They can be further subcategorized into fatty acyls, glycerolipids, sterol lipids, sphingolipids, and prenol lipids. The plasma levels of thirteen metabolites were negatively correlated with HAMA scores except 10-oxo-nonadecanoic acid. Based on the ROC curves, the fourteen lipid metabolites can be diagnostic biomarkers for PDA patients separately and the areas under the curve of the fourteen lipid metabolites ranged from 0.681 to 0.798. ConclusionsSignificantly lower plasma lipoproteins can be found in PDA patients. A panel of fourteen lipid metabolites were also significantly decreased and can be clinical biomarkers for the diagnosis of PDA patients.

Abstract 3442: Multimodal mapping of the immune landscape in human pancreatic cancer

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer related death in the US. Unfortunately, recent clinical trials using immunotherapy targeting the highly immunosuppressive tumor microenvironment have showed disappointing results, as there is no method to predict which patients will respond to therapy. More recently, the development of single cell technology has allowed for in-depth profiling at the cellular level using small amounts of tissue, raising the potential to develop precision medicine tools at time of endoscopic fine needle biopsy. Results: We performed single-cell RNA sequencing (scRNAseq) on endoscopic fine needle biopsies from 10 PDAC tumors at time of diagnostic endoscopic biopsy or 6 surgically resected tissues. We also sequenced 3 adjacent or normal pancreas tissues. We captured 8,521 cells from 3 surgical normal adjacent samples and 46,244 cells from PDAC tumors. Mapping of putative interactions between ligands and receptors demonstrated upregulation of key signaling pathways, including Hedgehog, NOTCH, and chemokine signaling within myeloid, epithelial, T, and NK cells. Differential expression analysis in cytotoxic CD8 T cells of PDA patients revealed increased expression of genes involved in T cell activation (GZMB, GZMA), exhaustion (GZMK, EOMES) as well as immune checkpoint pathway upregulation when compared to cytotoxic T cells in adjacent normal pancreatic tissue. Among the most significantly increased genes in CD8 T cells of PDAC tumor was the immune checkpoint TIGIT. Upon further analysis of the CD8 T cells, we found TIGIT was almost exclusively expressed in exhausted CD8 T cells, while other checkpoints such as PD-1 and LAG3 were equally distributed across effector and exhausted T cell populations. Interestingly, we were able to capture patient-specific heterogeneity of gene expression in T cells, suggesting the possibility of individualized T cell gene signatures present in PDAC tumors. We used mass cytometry and immunostaining to validate our transcript-based findings. Conclusion: Overall, we have successfully performed robust in-depth profiling using single-cell sequencing of PDAC tumors from fine needle biopsies. TIGIT, but not other immune checkpoints, correlates with T cells exhaustion in tumors, revealing an important biological function of this relatively understudied checkpoint. Analysis of our results identified patient-specific heterogeneity of key signaling pathways in different cell compartments of PDAC tumors that have to potential to be leveraged for precision medicine. Citation Format: Nina Steele, Eileen Carpenter, Samantha Kemp, Veerin Sirihorachai, Stephanie The, Lawrence Delrosario, Jenny Lazarus, El-ad Amir, Valerie Gunchick, Carlos Espinoza, Samantha Bell, Lindsey Harris, Valerie Irizarry-Negron, Dan Paglia, Justin Macchia, Fatima Lima, Angel Ka Yan Chu, Heather Schofield, Erik Jan Wamsteker, Richard Kwon, Allison Schulman, Anoop Prabhu, Ryan law, Arjun Sondhi, Katelyn Donahue, Hari Nathan, Clifford Cho, Michelle Anderson, Vaibhav Sahai, Costas Lyssiotis, Benjamin Allen, Arvind Rao, Weiping Zou, Filip Bednar, Timothy Frankel, Marina Pasca di Magliano. Multimodal mapping of the immune landscape in human pancreatic cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3442.