Human PC Research Articles

Abstract 1836: ONC201 shows synergistic effect with the androgen receptor AR-inhibitor darotulamide in prostate cancer models

Abstract Prostate cancer (PC) is the most frequently diagnosed cancer among men in the United States and is the 3rd cause of cancer mortality. Despite advances in the treatment and understanding of pathogenesis, patients with metastatic PC invariably progress to a lethal stage of castration-resistant prostate cancer (CRPC). Inhibition of androgen signaling remains crucial for the treatment of CRPC, but novel treatment strategies are urgently needed. ONC201 is a first-in-class, selective inhibitor of dopamine receptor D2 that upregulates death-receptor 5 and induces apoptosis. ONC201 induced apoptosis in PC cell lines and showed synergy with enzalutamide, docetaxel and everolimus (Lev A Mol Cancer Res 2018). Darolutamide (DARO) is a novel androgen receptor (AR) antagonist approved for treatment of patients with non-metastatic CRPC. DARO has higher affinity to AR and preclinical activity against enzalutamide-resistant PC cell lines including AR variants associated with enzalutamide agonism (Borgmann H Eur Urol 2018). We treated human PC cell lines (LNCAP [castration sensitive, AR wild-type], 22Rv1 [castration-resistant cell line that expresses AR splice variant AR-V7], DU145 [AR negative], PC3 [AR negative]) with DARO alone and combined with ONC201. Single-agent DARO decreased PC cell viability in Cell-Titer-Glo assays (IC50s of 10 and 693 nM for LNCAP and DU145, respectively). ONC201 showed strong synergism with DARO in LNCAP cells (combination indices < 1 at concentrations of 156, 312, 625 nM, and 1.25 µM of DARO with 2.5 μM ONC201). ONC201 induced robust apoptosis in 22Rv1 as measured by PARP cleavage, which was partially amplified by DARO. PARP cleavage was also observed in LNCAP cells with less intensity and not potentiated by DARO. ONC201 reduced the expression of phospho-AR (p-AR) in both 22Rv1 and LNCAP. The combination of ONC201 and DARO had a significant additive effect in reducing p-AR in 22Rv1, but not in LNCAP. ONC201 reduced PSA protein levels in LNCAP cells while DARO alone did not cause any significant change in PSA. Immunofluorescence experiments showed that DARO caused significant reduction of AR nuclear translocation in both 22rV1 and LNCAP cells. The combination with ONC201 potentiated this inhibition of AR translocation in both cell lines. ONC201 showed strong antiproliferative activity against PC cells lines independent of AR status. Combination of ONC201 with DARO demonstrated synergy in enzalutamide resistant 22Rv1 cells expressing AR-V7 with marked reduction of nuclear localization of AR. Our studies provide preclinical rationale for combination of ONC201 with DARO as a novel therapy of PC. Citation Format: Fabio Tavora, Lanlan Zhou, Ali Amin, Safran Howard, Navaraj Arunasalam, Andre de Souza, Anthony Mega, Dragan Golijanin, Wafik El-Deiry, Benedito Carneiro. ONC201 shows synergistic effect with the androgen receptor AR-inhibitor darotulamide in prostate cancer models [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1836.

A novel role for indoleamine 2,3-dioxygenase (IDO) in supporting the survival of long lived plasma cells (LLPC)

Abstract Protective antibodies against diseases such as measles and polio primarily come from bone marrow (BM) resident long lived plasma cells (LLPC) and are a key component of durable humoral immunity. As LLPC are not innately long lived a major question remains about how these cells maintain survival. Our lab has previously demonstrated that the T cell receptor, CD28, is essential for the survival and function of LLPC. Here we further demonstrate that when CD28 on LLPC ligates with CD80/CD86 on DC, it induces upregulation and activation of indoleamine 2,3-dioxygenase (IDO). IDO’s primary action is catabolizing tryptophan into kynurenine, a well-known ligand of the aryl hydrocarbon receptor (AhR). A nuclear transcription factor, AhR is known to modulate immune responses and is expressed in PC. Unexpectedly we observed that IDO KO mice have significantly less long lived antigen specific antibody titers and a decrease in LLPC specifically in the BM post immunization. Specific deletion of IDO+DC also significantly decreases LLPC in vivo. Furthermore, treatment with kynurenine in LLPC causes upregulation of CYP1A1, the downstream target of AhR. Moreover, we see an increase in CD28 expression with kynurenine treatment in murine and human PC. We propose a model where the interaction of CD28 on LLPC with CD80/CD86 on DC activates IDO, producing kynurenine, which activates AhR to further regulate CD28 expression on LLPC. These findings challenge the traditional paradigm of IDO as immunosuppressive and uncover a novel role in promoting immunity. Filling this gap in knowledge furthers our understanding of diseases that involve LLPC, but also has direct implications in future vaccine and therapeutic drug development. Support: T32 CA085183, R01CA121044

Interferon gamma inhibits CXCL8–CXCR2 axis mediated tumor-associated macrophages tumor trafficking and enhances anti-PD1 efficacy in pancreatic cancer

BackgroundPancreatic cancer (PC) is a common malignancy of the digestive system and is characterized by poor prognosis and early metastasis. Tumor immune escape plays an important role in PC progression....

Acinar transformed ductal cells exhibit differential mucin expression in a tamoxifen-induced pancreatic ductal adenocarcinoma mouse model.

ABSTRACTPancreatic cancer (PC) is acquired postnatally; to mimic this scenario, we developed an inducible KrasG12D; Ptf1a-CreER™ (iKC) mouse model, in which Kras is activated postnatally at week 16 upon tamoxifen (TAM) administration. Upon TAM treatment, iKC mice develop pancreatic intraepithelial neoplasia (PanIN) lesions and PC with metastasis at the fourth and fortieth weeks, respectively, and exhibited acinar-to-ductal metaplasia (ADM) and transdifferentiation. Kras activation upregulated the transcription factors Ncoa3, p-cJun and FoxM1, which in turn upregulated expression of transmembrane mucins (Muc1, Muc4 and Muc16) and secretory mucin (Muc5Ac). Interestingly, knockdown of KrasG12D in multiple PC cell lines resulted in downregulation of MUC1, MUC4, MUC5AC and MUC16. In addition, iKC mice exhibited ADM and transdifferentiation. Our results show that the iKC mouse more closely mimics human PC development and can be used to investigate pancreatic ductal adenocarcinoma (PDAC) biomarkers, early onset of PDAC, and ADM. The iKC model can also be used for preclinical strategies such as targeting mucin axis alone or in combination with neo-adjuvant, immunotherapeutic approaches and to monitor chemotherapy response.

Abstract C118: Transcriptome comparison of orthotopic and subcutaneous patient-derived xenografts

Abstract Background: Patient-derived xenograft (PDX) models are used for in vivo evaluation of oncology drugs. Tumors from cancer patients are implanted into immunodeficient mice either subcutaneously or orthotopically. Subcutaneous PDX models are more prevalently used because of easy operation and monitoring, albeit increasing evidence showed that the immune microenvironment in subcutaneous tissue is different from the original tract and is therefore less patient-like. By contrast, orthotopic PDX models have the capability to metastasize and match that of the donor patient well, but resulting in extensive inflammation. Previous studies showed that tumor location may affect immune response or result in different behavior of tumor expansion in mouse models [1-4]. There is yet insufficient study on the transcriptomic difference between subcutaneous and orthotopic PDX models. Method: Tumors for 10 PDX models—including four breast, four colorectal, and two liver cancers—were used to establish pairs of subcutaneous and orthotopic models. Tumors were dissected when reaching about 1000mm3, and subjected to transcriptomic profiling by RNAseq for gene expression comparison. Results: Using traditional alignment-based RNAseq analysis, we found hundreds of genes with different expression levels between subcutaneous and orthotopic models for the three cancers. Gene ontology analysis revealed that these differentially expressed genes are highly enriched in protein translation related biological processes and pathways, such as maturation of SSU-rRNA, cap-dependent translation initiation, and peptide chain elongation. Conclusion: Subcutaneous and orthotopic PDX models differ in transcriptomes that may affect protein translation and warrant further proteomics investigations. Reference: 1, Zhang, Yong, et al. “Real–Time GFP Intravital Imaging of the Differences in Cellular and Angiogenic Behavior of Subcutaneous and Orthotopic Nude–Mouse Models of Human PC–3 Prostate Cancer.” Journal of cellular biochemistry 117.11 (2016): 2546-2551. 2, Zhao, Xianda, et al. “Tumor location impacts immune response in mouse models of colon cancer.” Oncotarget 8.33 (2017): 54775. 3, Zhang, Yi, et al. “Establishment of a murine breast tumor model by subcutaneous or orthotopic implantation.” Oncology letters 15.5 (2018): 6233-6240. 4, Nakano, Kiyotaka, et al. “Difference in morphology and interactome profiles between orthotopic and subcutaneous gastric cancer xenograft models.” Journal of toxicologic pathology 31.4 (2018): 293-300. Citation Format: xiaoqian jiang, henry li, sheng guo. Transcriptome comparison of orthotopic and subcutaneous patient-derived xenografts [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr C118. doi:10.1158/1535-7163.TARG-19-C118

A Smart Home Monitoring System for Abnormal Human Activity Detection using CNN

Demonstrating human practices and movement designs for acknowledgment or location of exceptional occasion has pulled in noteworthy research enthusiasm for late years. Differing strategies that are flourish for structure smart vision frameworks went for scene comprehension and making right semantic derivation from the watched elements of moving targets. Most applications are in reconnaissance, video content recovery, and human PC interfaces. In this propose a novel strategy for irregular human action recognition in jam-packed scenes/Home. In particular, as opposed to recognizing or fragmenting people, we formulated a productive technique, called a movement impact map, for speaking to human exercises. The key element of the proposed movement impact guide is that it viably mirrors the movement qualities of the development speed, development bearing, and size of the items or subjects and their communications inside an edge succession. In this propose System developing using CNN.

2019P - Protein tyrosine phosphatase non-receptor type 3 (PTPN3) could be a new therapeutic target for pancreatic cancer

BackgroundPancreatic cancer (PC) is a refractory disease and we have not an effective therapeutic strategy yet. Therefore, the development of a new therapeutic procedure is urgently required. Recently, protein tyrosine phosphatase (PTP) has been attracted as a new therapeutic target for some types of cancers. However, the biological significance of PTP in PC has never been elucidated yet. In the present study, to develop effective therapeutic strategy for refractory PC, the biological significance of PTP non-receptor type 3 (PTPN3) in PC is investigated. MethodsThree pancreatic ductal adenocarcinoma cell (PDAC) lines were used as target cells. Inhibition or overexpression of PTPN3 was performed using PTPN3 siRNA/shRNA and plasmid, respectively. Protein expression was analyzed by western blot and immunostaining. Proliferation assay was performed by MTT assay. Migration and invasion were estimated by time-lapse imaging and matrigel invasion assay. Mice xenograft experiments were performed using BALB/c nude mice. Twenty-three surgically resected human PC specimens were used for immunostaining. Results1) PTPN3 is highly expressed in PC cells compared to that in normal pancreatic duct cells by immunostaining. 2) Inhibition of PTPN3 significantly decreased migration and invasion in PDAC through inhibition of epithelial mesenchymal transition (EMT) and matrix metalloproteinase (MMP)-2 expression. 3) PTPN3 inhibition significantly decreased proliferation in vitro in PDAC. 4) PTPN3 overexpression led to increased proliferation, migration and invasion in PDAC. 5) Tumor volume in mice injected with PTPN3-inhibited PDAC was significantly lower than that in control mice. The expressions of Ki-67 and VEGF in PTPN3-inhibited PDAC were lower than those in control. 6) Signaling from PTPN3 was through PI3K and MAPK signaling pathways. 7) When intensity of PTPN3 expression was divided into 2 groups; weak and strong, intensity of PTPN3 expression was positively correlated with tumor size (T factor) and clinical staging. ConclusionsThese results suggest that PTPN3 contributes to the induction of malignant phenotypes of pancreatic cancer and that PTPN3 could be a new effective and specific therapeutic target for PC. Legal entity responsible for the studyKyushu University Hospital, Cancer Therapy and Research. FundingHas not received any funding. DisclosureAll authors have declared no conflicts of interest.

Hand Gesture Recognition System Using Deep Learning

Hand motion acknowledgment is a characteristic method for human PC association and a zone of dynamic research in PC vision and AI. This is a zone with a wide range of conceivable applications, giving clients an easier and increasingly normal approach to speak with robots/frameworks interfaces, without the requirement for additional gadgets. Along these lines, the essential objective of signal acknowledgment explore connected to Human-Computer Interaction (HCI) is to make frameworks, which can distinguish explicit human motions and use them to pass on data or controlling gadgets. For that, vision-based hand signal interfaces require quick and incredibly strong hand discovery, and motion acknowledgment continuously. This paper introduces an answer, sufficiently nonexclusive, with the assistance of deep learning, permitting its application in a wide scope of human-PC interfaces, for ongoing motion acknowledgment. Investigations did demonstrated that the framework had the capacity to accomplish a precision of 99.4% as far as hand act acknowledgment and a normal exactness of 93.72% as far as unique signal acknowledgment.

Abstract 2165: Self-microemulsifying drug delivery systems (SMEDDS) containing novel compounds in combination with gemcitabine: Bioavailability enhancement and synergistic anticancer efficacy studies for pancreatic cancer management

Abstract Pancreatic cancer (PC) is the third leading cause of deaths in the United States claiming approximately 45,000 lives every year and is predicted to be the second cause of cancer related deaths by 2020. Late diagnosis, poor prognosis, limited success and significant side effects with existing anticancer drugs has resulted in such high mortality rates (5 year survival rate is less than 8%). These limitations have encouraged scientists to develop chemopreventive strategies and combination drug alternatives that can serve as better therapeutics. In our previous studies, we have reported the synergistic chemoprevention potential (formulation patent pending) of lipid nanocarriers (SMEDDS) of Loratadine (LOR), H1-receptor antagonist drug; with free Sulforaphane (SFN), a neutraceutical compound from broccoli when administered at low doses in human PC cell lines (MIA PaCa-2 and Panc-1). To confirm the hypothesis that enhanced efficacy using the SMEDDS formulation results from bioavailability enhancement, preclinical in vivo pharmacokinetic studies were performed in Sprague Dawley rats. The results confirmed that the bioavailability and Cmax of LOR increased 5-fold (2203.2 ± 293.7 ng.h/mL for plain LOR and 10266.1 ± 250.0 ng.h/mL for LOR SMEDDS) and 4-fold (120.9 ± 10.5ng/mL for plain LOR and 503.2 ± 5.8ng/mL for LOR SMEDDS) when administered as SMEDDS compared to plain drug. This corroborated the significant enhancement in bioavailbility with SMEDDS formulation. This promising results in bioenhancement and chemoprevention activity led to an hypothesis to assess the potential of LOR SMEDDS-SFN combination as an adjuvant therapy with classical anticancer drugs like Gemcitabine (GEM) to give synergistic efficacy enhancement in PC treatment. For this, IC50 concentrations were calculated for various treatment groups [GEM alone or in addition to the LOR free drug or as SMEDDS with or without SFN] using MTS viability assay on human PC cell lines in accordance with published protocol over an incubation time of 72 h. The LOR SMEDDS-SFN-GEM combination revealed 2-fold and 2.5-fold enhancement in anticancer activity compared to free drug combination and GEM alone respectively. This confirmed the synergistic anticancer potential of the free drug combination followed by further activity potentiation using SMEDDS formulation. In conclusion, a novel SMEDDS of LOR and free SFN was invented (provisional patent applied) to successfully elicit PC chemoprevention and adjuvant treatment in combination with GEM. We anticipate that, this adjuvant therapy will lead to relatively low doses of anticancer drugs and thereby can reduce the side effects with enhanced efficacy. Citation Format: Sunil Prabhu, Preshita Desai, Naga Jyothi Thumma, Jeffrey Wang. Self-microemulsifying drug delivery systems (SMEDDS) containing novel compounds in combination with gemcitabine: Bioavailability enhancement and synergistic anticancer efficacy studies for pancreatic cancer management [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2165.

Hand recognition in gaming

Hand motion acknowledgment gives a savvy and characteristic method for human PC collaboration (HCI). It has applications running from customer hardware to control of therapeutic recovery (for example cell phone). So as to recognize hand motions, different sorts of detecting methods are used to get signals for example acknowledgment. Increasing speed base and electromyogram-based strategies are two research branches in the field of hand signal example acknowledgment. Quickening based (ACC-based) motion control is typically contemplated as a valuable connection methodology. It is appropriate to recognize perceptible, bigger scale signals with various hand directions of lower arm developments. With ACC-based strategies some unobtrusive finger or hand development might be overlooked while electromyogram-based (EMG-based) motion acknowledgment systems use multi-channel EMG signals which contain rich data about hand motions of different size scales. Because of certain issues natural in the EMG estimations, including the distinguishableness and reproducibility of estimation, the size of discriminable hand signal set is as yet restricted to 4-8 classes. So as to understand a characteristic and powerful signal based HCI framework, the determination of info hand motions that are well discriminable from one another is of essential significance. Thinking about the integral highlights of ACC-and EMG-estimations, we accept that their mix will expand the quantity of discriminable hand, wrist and lower arm signals and the precision of the acknowledgment framework. This paper depicts In IP Gaming we are proposing a framework wherein without utilizing sensors and Devices, we are recognizing the hand and signal with Simple Web camera and playing out the Image Processing system in which utilizing those motion, we can play game on reassure. In Image Process Gaming, the movements are identified through a web camera. These pictures are then passed for the picture handling. The procedures utilized for picture handling are hand motion identification, edge recognition, thresholding, form location. Utilizing OpenCV, which gives a library accumulation of capacities for various picture handling systems, these info pictures can be prepared and relating key strokes will be created. As of late, PC vision improvement has extraordinary headways and our everyday life undertakings are fragmented without utilizing PCs. The real info gadgets like Keyboard and mouse are accustomed to communicating with PCs. Among the different communication systems utilization of hands as an information is an appealing technique for building up regular Human Computer Interaction. By utilizing Hand motions client can convey more data in less Timespan. So for improving the interface among clients and PCs human PCs collaboration (HCI) innovation has incredible use.

Abstract A050: Integrated system-level analyses define an androgen receptor variant oncogenic network for advanced prostate cancer

Abstract Unbiased high-throughput approaches provide a global view of the massive genetic and transcriptomic changes that occur in and potentially drive cancer. Such approaches permit the discovery of novel networks that can serve as prognostic markers and therapeutic targets. We performed a systems-biology approach to identify genes linked to the androgen receptor (AR) variant AR-V7 in castration-resistant prostate cancer (CRPC). AR variants are C-terminal truncated, constitutively active, and play key transcriptional regulatory roles resulting in treatment resistance and disease progression. These AR variants lack the C-terminal ligand binding domain, which is the target of all approved drugs against AR, and consequently there is considerable interest in novel therapeutic strategies that are effective against AR variant-expressing prostate tumors. Directly targeting AR variants is challenging, and thus an attractive alternative approach is to identify downstream AR-V7 hub genes that not only drive cancer progression but also feedback to enhance AR-V7 activity. Thus, our goals were to: 1) define clinically relevant gene sets or modules in human PC progression using unbiased gene expression meta-analysis of PC patient samples by Weighted Gene-Co-expression Network Analysis (WGCNA); 2) map the AR-V7 functional interactome using a unique high-throughput synthetic genetic array (SGA) screening in the yeast S. pombe; and 3) identify AR-V7 regulated genes in PC cells. Integration of these approaches resulted in the identification of seven genes that are upregulated in human PC (as determined by WGCNA), functionally interact with AR-V7 (as determined by SGA), and are regulated by AR-V7. This gene set encoded select proteins previously linked to PC, such as members of the cyclin and topoisomerase families, but also proteins with no prior association with AR-V7 or PC. The seven genes comprised a signature that correlated with patient Gleason score and predicted disease free-survival and overall survival in large independent PC patient cohorts. Individual depletion of the expression of these genes decreased ligand-independent AR transcriptional activity concomitant with reduced CRPC cell proliferation. Because CRPC cells were highly dependent on these inter-related genes, combination therapy targeting two of the seven genes led to potent inhibition of CRPC cell proliferation. In contrast, the combination of these agents had little to no effect on nontumorigenic prostate epithelial cells or AR-null PC cells. In summary, we utilized an unbiased and novel gene discovery strategy that combined sophisticated bioinformatics and experimental approaches. This multifaceted approach resulted in the identification of disease-relevant genes that were regulated by AR-V7 and that reciprocally enhanced AR-V7 oncogenic activity. This novel gene set has a prognostic value and provides a rationale for combinatorial CRPC therapy. Citation Format: Fiorella Magani, Eric Bray, Maria Julia Martinez, Valeria Copello, Ning Zhao, Stephanie Peacock, David Wiley, Gennaro D’urso, Kerry L. Burnstein. Integrated system-level analyses define an androgen receptor variant oncogenic network for advanced prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Prostate Cancer: Advances in Basic, Translational, and Clinical Research; 2017 Dec 2-5; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(16 Suppl):Abstract nr A050.

Abstract 1314: A prostate cancer androgen receptor variant gene network with prognostic and therapeutic value identified using integrated systems analyses

Abstract Constitutively active androgen receptor (AR) splice variants such as AR-V7 represent a well-established mechanism of disease progression to castration-resistant prostate cancer (CRPC), the incurable form of PC. These AR variants lack the carboxyl-terminal ligand binding domain, the target of all approved drugs against AR. Consequently, a significant challenge is to identify novel therapeutic strategies that are effective against AR variants-expressing prostate tumors without requiring direct targeting of these variants, which is challenging due in part to the unstructured nature of the AR amino terminus. We used a multi-pronged, unbiased systems biology approach to identify downstream AR-V7-regulated hub genes that both drive cancer progression and feedback to enhance AR-V7 activity. Such genes likely encode prognostic markers as well as potential therapeutic targets acting within the AR-V7 network. Our systems biology approach utilized: 1) clinically relevant gene sets upregulated during human PC progression obtained by Weighted Gene-Co-expression Network Analysis (WGCNA); 2) an AR-V7 functional interactome from a high-throughput synthetic genetic array (SGA) screening in the yeast, Schizosaccharomyces pombe; and 3) PC cell AR-V7 transcriptome. We identified seven genes that were upregulated in human PC, functionally interacted with AR-V7 and were AR-V7 targets in PC. This gene set, composed of cell cycle and mitosis-regulating genes, not only encoded select proteins previously linked to PC, such as Cyclin B1 and Topoisomerase 2A (CCNB1 and TOP2A), but also proteins with little or no prior association with AR-V7 or PC. The seven genes comprised a signature that strongly correlated with patient Gleason score and predicted disease free-survival in large independent PC patient cohorts. In contrast, this gene set was not associated with survival metrics in other human cancers. Individual depletion of the expression of these genes decreased ligand-independent AR transcriptional activity and reduced CRPC cell proliferation. Furthermore, individual knock down of the members of the gene set affected the expression of the other members of the set indicative of the interrelationship of these genes. Consistent with CRPC dependence on these interrelated mitotic genes, combinatorial targeting of two of its members: TOP2A and CCNB1, led to a synergistic and selective inhibition of AR-driven CRPC cell proliferation but had no effect on non-tumorigenic prostate epithelial cells. This unbiased and novel gene discovery strategy, which is broadly applicable to other cancers, identified clinically-relevant interacting gene hubs forming the basis for both prognostic use and rational, combinatorial therapy for CRPC. Citation Format: Maria J. Martinez, Fiorella Magani, Eric R. Bray, Valeria Copello, Ning Zhao, Stephanie Peacock, David J. Wiley, Gennaro D'Urso, Kerry L. Burnstein. A prostate cancer androgen receptor variant gene network with prognostic and therapeutic value identified using integrated systems analyses [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1314.

Abstract 4062: Novel synthetic peptide targets CD206 receptor promoting the repolarization of prostate cancer (PCa) TAMs and increase the efficacy of conventional prostate cancer therapeutic

Abstract Background: Tumor associated M-ϕs (TAMs) have a complex etiology as a result of soluble factors secreted by prostate tumor cells, creating an immunosuppressive micro-environment (M-E) for the progression of late stage PC that has metastasized to the bone. The result is an M2-like polarization that expresses a number of M2-TAM specific markers, including CD206. Therapeutic targeting of TAMs has proven to be a challenge, due partly to a lack of understanding of the function of this biomarker and the bi-directional effects of TAMs on tumor cells. Our objective is to characterize the CD206 receptor in PC patients, and to test the potential of a novel engineered CD206 targeted peptide (T-P) to reverse the immune-suppressive M-Env in late stage PC. Methods: The TGCA Prostate Dataset was used for patient data and R software was used for informatics analysis. Small-angle X-ray scattering (SAXS) was used to generate the NMR structure of CD206. Peptide binding to the naïve CD206 receptor was determined in silico using PIPER, and was validated by Surface Plasmon Resonance (SPR). In vivo efficacy of CD206 T-P was determined in mice inoculated with PC-3 intratibially. Mice were treated with peptide alone, daily or twice a week at 10, 20, and 50mg/Kg, as a monotherapy or in combination with 20mg/Kg Docetaxel (DTX) over a 21day period. CD86 (M1) and CD206/CD163 (M2-TAM) markers and pro-inflammatory cytokines were determined by FACS. Bone osteolysis was determined by IHC of the entire bone, and bone turnover was determined by IHC analysis of RANKL. Results: TCGA data showed that over-expression of CD206 in Human PC is significantly associated with poor survival. In silico analyses showed the T-P exhibited strong affinity towards CD206 and was validated by SPR (Kd =3.57 µM). In bone-metastatic xenograft models of CRPC (PC-3), the flux differential of relative fluorescence in IVIS imaging showed peptide alone (at 20mg/kg qD sQ) decreased fluorescence by 64.36% compared with the untreated group. FACS analyses of M1/M2 markers CD86/CD206 in mice treated with peptide showed a 33.87 fold decrease in CD206 (M2-TAM) expression and a 7.6 fold increase in CD86 (M1-TAM) expression, compared with only a 4.4 fold decrease in CD206 expression and 0.44 fold decrease in CD86 expression in DTX treated mice. Stem cell marker CD133 showed a 4.7 fold decrease expression compared to the untreated group further suggesting that TAM repolarization reversed the immunosuppressive M-E. X-ray of tumors in bone suggests a reduction of osteolysis and was confirmed by IHC staining and analysis of whole bone showing fibrous and inflammatory remodeling patterns of tumor regression in the peptide only group. Conclusion: These results suggest that the novel engineered CD206 T-P repolarizes M2-TAM phenotype to M1-TAM which reverses the immunosuppressive cancer M-E in late stage PC. Citation Format: Ahmad B. Salam, Benjamin Adu-Addai, Huixian Lin, Anghesom Ghebremedhin, Jason White, Ruksana Amin, Balasubramanyam Karanam, George Martin, Charles Garvin, Henry Lopez, Jesse M. Jaynes, Clayton Yates. Novel synthetic peptide targets CD206 receptor promoting the repolarization of prostate cancer (PCa) TAMs and increase the efficacy of conventional prostate cancer therapeutic [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4062.

Abstract 1097: The Phlpp2 phosphatase protects Myc and is a target for the prevention of prostate cancer metastasis

Abstract Prostate cancer (PC) is one of the most prevalent cancers among men, causing almost 30,000 deaths per year in the US alone. Death caused by PC is mainly due to metastasis, and the 5-year survival rate of metastatic PC is only 28%. In contrast, the 5-year survival rate of PC confined to the prostate is nearly 99%. In contrast to primary PC tumors, metastatic tumors commonly present with targeted, bi-allelic mutations of Pten and Tp53 tumor suppressor genes. Whereas 49% of metastatic PC tumor samples have Pten/Tp53 co-deletions, these same co-deletions arise in only 10% of primary tumors. Because of the scarcity of human samples from metastatic disease, our lab has developed a genetically engineered mouse model (GEMM), in which Pten/Trp53 co-deletion using lentiviral plasmids gives rise to metastatic PC. As a result of the propensity of these mice to quickly develop metastatic PC, the model is termed RapidCaP. In the setting of our model for metastatic PC, we sought to investigate other genes which could be regulating the switch from a relatively benign primary PC to a highly lethal metastatic PC. Interestingly, most candidate tumor suppressor genes in PC are part of large recurrent hemizygous deletions, such as the common chromosome 16q-deletion, which involves the AKT-suppressing phosphatase, Phlpp2. Using our RapidCaP model, we find evidence that complete loss of Phlpp2 in the setting of Pten/Tp53 co-deletion paradoxically blocks prostate tumor growth and metastasis burden in RapidCaP mice. In fact, when Phlpp2 is deleted using a lentiviral vector, only 7% of the mice have PC tumors with metastasis compared to 60% in Pten/Tp53 co-deletion alone. Furthermore, we show that the Phlpp2 phosphatase activates Myc, a key driver of PC metastasis, by regulating Myc stability. Phlpp2 deletion in embryonic fibroblasts (MEFs) derived from RapidCaP mice does not affect Myc transcript levels by qPCR, but decreases Myc protein expression by over 75% as measured by Western blot. Using the same MEFs, we further show that the half-life of the Myc protein decreases from 19.3 minutes to 12.7 minutes upon deletion of Phlpp2 using lentiviral vectors. We propose that Phlpp2 dephosphorylates the threonine 58 (T58) site of Myc, thus directly increasing its stability. Importantly, we show that small molecule inhibitors of the Phlpp2 phosphatase can suppress Myc and cause cell death. Using the MEFs derived from the RapidCaP mice, Myc concentration decreases to an undetectable level in a dose-dependent manner following treatment with a Phlpp2 inhibitor between the concentrations of 0-200 υM, which coincides with a dose-dependent increase in cell death as measured by PI staining. A similar trend is seen in the PC3 cell line derived from human PC. In sum, our findings reveal that the frequent hemizygous deletions on chromosome 16q present a possible druggable vulnerability for targeting the Myc protein in metastatic PC with Phlpp2 phosphatase inhibitors. Citation Format: Vincent D'Andrea, Dawid Nowak, Kaitlin Watrud, Alexandra Ambrico, Irene Casanova-Salas, Lloyd Trotman. The Phlpp2 phosphatase protects Myc and is a target for the prevention of prostate cancer metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1097.

Abstract 2323: Involvement of glutamine in tolerance of human pancreatic cancer cell to inhibition of fatty acid synthesis

Abstract Pancreatic cancer (PC) is the fifth leading cause of cancer-related mortality in Japan. Although the standard chemotherapy regimens, FOLFIRINOX (fluorouracil, leucovorin, irinotecan and oxaliplatin), gemcitabine and nab-paclitaxel, are widely used to treat patients with advanced pancreatic cancer, their antitumor effects (progression and survival rates) are less potent against this cancer type than other solid tumors. Previously, we found that inhibition of acetyl-CoA carboxylase (ACC), a major rate-controlling enzyme of fatty acid synthetic pathway, suppressed the proliferation of PC cells. However, in this study, we found the tolerance to the inhibition in a part of PC cells. Therefore, we investigated the mechanism of the tolerance to ACC inhibition in PC cells. The human PC cell lines, AsPC-1, BxPC-3 and PANC-1 were obtained from the American Type Culture Collection. Live and apoptotic cell numbers were determined using the MUSE Annexin V and Dead Cell Kit according to the manufacturer's instructions. PANC-1 cells showed the tolerance to inhibition of fatty acid synthesis by ACC inhibitor, 5-(tetradecyloxy)-2-furoic acid (TOFA), whereas TOFA induced apoptosis in AsPC-1 and BxPC-3 cells. In addition, PANC-1 did not show autophagy even under the condition of fatty acid starvation. Next, we investigated whether PANC-1 has the tolerance to nutrient starvation in nutrient-depletion medium, Earle's Balanced Salt Solution (EBSS) and serum-free medium respectively. PANC-1 showed the survival even in such mediums, whereas AsPC-1 and BxPC-3 almost dead. This result suggests that the tolerance of fatty acid starvation of PANC-1 are related to that of nutrient starvation. Interestingly, proliferation of PANC-1 cultured in glutamine-depletion medium dramatically was suppressed, even though low glucose medium did not affect. BPTES, a selective inhibitor of glutaminase, also suppressed the proliferation and induced apoptosis in PANC-1, whereas there are little effects of 2-dexy-glucose, inhibitor of glycolysis, on them. These results suggest that the TCA cycle plays an important role on the tolerance to inhibition of fatty acid synthesis and nutrient starvation of PANC-1, and can be novel therapeutic target of PC cells which have the tolerance to nutrient starvation like PANC-1. Citation Format: Koji Nishi, Mina Suzuki, Noriko Yamamoto, Yumiko Iwase, Nagahiko Yumita. Involvement of glutamine in tolerance of human pancreatic cancer cell to inhibition of fatty acid synthesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2323.

A novel audio based human interaction proof for visually challenged users

CAPTCHAs are strategies to recognize human clients and PC programs naturally. CAPTCHAs shield different sorts of online administrations from beast compel assaults and foreswearing of administration via programmed PC programs. Most CAPTCHAs comprise of mages with misshaped content. Shockingly, visual CAPTCHAs constrain access to the a huge number of outwardly hindered individuals utilizing the Web. Sound CAPTCHAs were made to fathom this openness issue. However the presently accessible sound CAPTCHAs have been broken with differing achievement, utilizing the shortcoming in the techniques utilized. Our system, presents the user with an interface that plays a song using instrumental music (nonvocal) randomly selected from some language of users choice. The user is then asked to kind the music composer and then the device estimates whether it is a human or no longer by means of analyzing the response. A person look at turned into conducted to research the overall performance of our proposed mechanism.

Возможности использования «Натрия фторида, 18F» для диагностики опухолей в костях методом позитронной эмиссионной томографии (экспериментальное исследование)

Purpose: To study the potential of using radiopharmaceutical “Sodium Fluoride 18F” in order to reveal foci of malignant tumors in animals’ bones by positron emission tomography (PET).
 Material and methods: Two experimental models (human prostate adenocarcinoma PC 3 and Pliss lymphosarcoma) were used. Tumor cell PC 3 suspensions were injected under a hip periosteum of the BALB/c nude line mice or outbred rats (Pliss lymphosarcoma). Radiopharmaceutical was injected into a tail vein of animals with developed tumors. Then PET scanning was performed for these animals.
 Results: Effective half-life of radiopharmaceutical from blood was 9.45 min, from skeleton bones - 77.5 min and tumors – 99 min. The hyperfixation level of radiopharmaceutical in the tumor was sufficient to image tumors by PET. In areas of the tumor from cells of prostate adenocarcinoma PC 3 and Pliss lymphosarcoma the uptake rate of radiopharmaceutical was quite high during an hour that resulted in a high contrast image of tumors in animals’ bones. T/NT ratio tumor/bone in rats with Pliss lymphosarcoma was increased in the first 20 min after administration of the radiopharmaceutical, and then slowly decreased. Nevertheless, in 40 minutes after injection of the radiopharmaceutical T/NT ratio tumor/bone remained quite high – more then 1.7, and T/NT ratio tumor/muscle exceeded to 9.0.
 Conclusion: The uptake level and fixation time of the radiopharmaceutical in tumor foci, as well as low activity value and its rapid excretion from intact tissues show the expediency of the use of “Sodium Fluoride 18F” to diagnose intraosseous foci of malignant diseases.

Abstract 1159: Targeting novel mucin synthesis pathway for pancreatic cancer treatment

Abstract Pancreatic cancer (PC) is most fatal and is the fourth leading cause of cancer related deaths in USA with a least five year survival of <7%. Although several overexpressed mucin subtypes (MUC1, MUC4, MUC5AC, and MUC16) that impede drug delivery to PC have been targeted with limited results, core enzymes involved in mucin biosynthesis have yet to be evaluated as potential targets. The core 2 β 1,6 N-acetylglucosaminyltransferase (GCNT3/C2GNT) plays a significant role in mucin glycan biosynthesis. We analyzed GCNT3 expression on 180 specimens from individuals with PC (90) or matched controls (90). Patients with low GCNT3 expression survived longer than patients with high GCNT3 expression (median survival: 17.5 vs. 10.5 months, p=0.036). We observed a significant increase in GCNT3 mRNA expression (103.16 folds; p<0.0001) in correlation with increased mucins subtypes (p<0.05) in the pancreatic tumors from genetically engineered mouse (GEM) compared with pancreata from wild-type mice, as determined by Next-generation Sequencing/RNAseq analysis. In in silico studies, blind docking simulations revealed that a novel drug talniflumate binds to GCNT3 with a better docking affinity of -8.3 kcal/mol. We synthesized talniflumate and determined quality/purity using NMR, HRMS, and HPLC for in vitro and in vivo studies. Pancreata from 6-week-old Kras GEM (N=6/group) treated with talniflumate showed a significant decrease in GCNT3 and mucin expression in PanIN lesions. In human PC MiaPaCa cell line, talniflumate reduced GCNT3 expression and the number of mucin-positive cells (38% p<0.0001). Further, PC cells (BxPc3 and Panc1) were treated with talniflumate for 24 hrs for mucin disruption followed by radioactive 3H gemcitabine treatment for 2 hrs. The cells treated with talniflumate showed 40% (p<0.03) and 50% (p<0.025) increased 3H gemcitabine uptake in BxPc3 and Panc1 cells compared to respective controls. GCNT3 CRISPR knock out PC cells and talniflumate treated PC cells showed a reduction in cell proliferation and significantly reduced spheroid formation (p<0.05). In in vivo, we did not observe any overt toxicity upon talniflumate treatment as determined by liver enzyme toxicity and body weight gain (N=5 mice/group). BxPC3 cells implanted nude mice (N=5 mice/group) treated with talniflumate or GCNT3 siRNA showed a marked reduction in xenograft tumors compared with untreated controls. Both talniflumate and GCNT3 siRNA significantly inhibited xenograft tumor weight (48% p<0.002 with siRNA, 63% p<0.0001 with talniflumate) and tumor volume (75% p<0.0001 with both siRNA and talniflumate) and GCNT3 enzyme activity (28% p<0.03). Transcriptome analysis of xenograft tumors (N=3/group) treated with GCNT3 SiRNA or talniflumate showed reduced GCNT3 and top hits related to mucin expression. These results justify the use of talniflumate for mucin synthesis disruption, thereby enhancing the uptake of gemcitabine and thereby efficacy for PC treatment. Citation Format: Altaf Mohammed, Naveena B. Janakiram, Venkateshwar Madka, Gaurav Kumar, Scott Edgar, Gopal Pathuri, Taylor Bryant, Yuting Zhang, Hariprasad Gali, Yan Daniel Zhao, Stan Lightfoot, Chinthalapally V. Rao. Targeting novel mucin synthesis pathway for pancreatic cancer treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1159. doi:10.1158/1538-7445.AM2017-1159

Abstract 1182: A novel curcumin derivative inhibits active ras and its downstream pathway in pancreatic cancer

Abstract Pancreatic Cancer’s (PC’s) 5-year survival rate of only 6.7% indicate the need to improve treatment modalities. Despite decades of research, current chemotherapy and radiation therapy regimens offer minimal or no help. It is critical to develop new agents for the effective management of PC. Given that Kras mutations initiate and maintain PC, inhibition of this pathway is widely considered a therapeutic target of exceptional importance. Our laboratory is currently exploring the chemotherapeutic efficacy of a novel Chemically-Modified Curcumin (CMC2.24) as a potential chemotherapeutic agent for PC. Preliminary studies have shown that CMC2.24 has higher bioavailability than curcumin, as shown in pharmacokinetic studies in rats, and inhibits PC growth in vitro and in vivo. However, the exact mechanism on how CMC2.24 reduces cell growth remains unidentified. The objective of this work was to determine the mechanism of CMC2.24 in PC. Using human PC MIA PaCa-2 and Panc-1 cell lines and pancreatic acinar explants from Kras mutant mice, we explored the effects of CMC2.24 on Ras activation, ERK phosphorylation, mitochondrial reactive oxygen species, mitochondrial ATP production, and intrinsic apoptosis. In human PC MIA PaCa-2 cells CMC2.24 inhibited Ras activation by 90% (p<0.05). This was confirmed in primary acinar explants isolated from Kras mutant mice in which CMC treatment reduced Ras activation by 70%, compared to control. Furthermore, CMC 2.24 treatment reduced the phosphorylation MEK, ERK and c-Raf, down-stream pathway of Ras, both in vitro and in vivo. The effect of CMC2.24 on ERK phosphorylation was confirmed by Immunofluorescence, showing a significant decrease in ERK phosphorylation (47.8%; p<0.05). Moreover, CMC2.24 treatment enhanced the levels of superoxide anion in mitochondria by 200% (p<0.02), decreased ATP levels in a concentration-dependent manner (p<0.05), and induced intrinsic apoptosis, as shown by the increase in caspase 9 and Parp cleavage downstream of cytochrome C cytosolic release. In conclusion, our results indicate that the Ras pathway is a key molecular target for CMC2.24 and that CMC induces apoptosis in PC cells through the intrinsic pathway. This research was supported by the Stony Brook Cancer Center and a URECA Summer Grant. Citation Format: Naveen Mallangada, Gerardo G. Mackenzie. A novel curcumin derivative inhibits active ras and its downstream pathway in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1182. doi:10.1158/1538-7445.AM2017-1182

DNA aptamer selected for specific recognition of prostate cancer cells and clinical tissues

Prostate cancer is the most common malignancy in men lack of efficient early diagnosis and therapeutics, calling for effective molecular probes. Herein, we performed cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX) to obtain specific recognition of human prostate cancer cells PC–3M. Four aptamers were successfully obtained that can bind to target cells with high affinity and specificity. A 51-nt truncated sequence named Xq-2-C1 was identified after further elaborative analysis on the secondary structure. More importantly, the achieved aptamer Xq-2-C1 not only demonstrated excellent specific to target cells, but also revealed specific recognition to clinical prostate cancer tissue. The tissue imaging results showed that Xq-2-C1 had better recognition ratio for clinical prostate cancer tissue samples (85%) compared to the random sequence (9%). These results demonstrate that these newly generated aptamers would furnish potential applications in the early diagnosis and clinical treatment of prostate cancer.