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Annexin A1 (AnxA1), a phospholipid-binding protein and regulator of glucocorticoid-induced inflammatory signaling, has implications in cancer. Here, a role for AnxA1 in prostate adenocarcinoma was determined using primary cultures and a tumor cell line (cE1), all derived from the conditional Pten deletion mouse model of prostate cancer. AnxA1 secretion by prostate-derived cancer-associated fibroblasts (CAF) was significantly higher than by normal prostate fibroblasts (NPF). Prostate tumor cells were sorted to enrich for epithelial subpopulations based on nonhematopoietic lineage, high SCA-1, and high or medium levels of CD49f. Compared with controls, AnxA1 enhanced stem cell–like properties in high- and medium-expression subpopulations of sorted cE1 and primary cells, in vitro, through formation of greater number of spheroids with increased complexity, and in vivo, through generation of more, larger, and histologically complex glandular structures, along with increased expression of p63, a basal/progenitor marker. The differentiated medium-expression subpopulations from cE1 and primary cells were most susceptible to gain stem cell–like properties as shown by increased spheroid and glandular formation. Further supporting this increased plasticity, AnxA1 was shown to regulate epithelial-to-mesenchymal transition in cE1 cells. These results suggest that CAF-secreted AnxA1 contributes to tumor stem cell dynamics via two separate but complementary pathways: induction of a dedifferentiation process leading to generation of stem-like cells from a subpopulation of cancer epithelial cells and stimulation of proliferation and differentiation of the cancer stem-like cells.Implications: AnxA1 participates in a paradigm in which malignant prostate epithelial cells that are not cancer stem cells are induced to gain cancer stem cell–like properties. Mol Cancer Res; 12(4); 607–21. ©2014 AACR.

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PDF file - 819K, List of primer sequences for semi-quantitative and quantitative PCR

PDF file - 504K, Response of xenografts of human prostate cancer specimens to IGF-1R inhibition as a function of NRP2 expression

PDF file - 454K, Role of JNK/c-Jun in regulating NRP2 expression in prostate carcinoma cells

PDF file - 1.8MB, Evaluation of insulin signaling in prostate tumors and cell lines as a function of NRP2 expression and role of VEGF in regulation of Bmi-1

We show that the VEGF receptor neuropilin-2 (NRP2) is associated with high-grade, PTEN-null prostate cancer and that its expression in tumor cells is induced by PTEN loss as a consequence of c-Jun activation. VEGF/NRP2 signaling represses insulin-like growth factor-1 receptor (IGF-IR) expression and signaling, and the mechanism involves Bmi-1–mediated transcriptional repression of the IGF-IR. This mechanism has significant functional and therapeutic implications that were evaluated. IGF-IR expression positively correlates with PTEN and inversely correlates with NRP2 in prostate tumors. NRP2 is a robust biomarker for predicting response to IGF-IR therapy because prostate carcinomas that express NRP2 exhibit low levels of IGF-IR. Conversely, targeting NRP2 is only modestly effective because NRP2 inhibition induces compensatory IGF-IR signaling. Inhibition of both NRP2 and IGF-IR, however, completely blocks tumor growth in vivo.Significance: These results identify a causal role for NRP2 and VEGF/NRP2 signaling in the behavior of aggressive prostate cancers by a mechanism that involves regulation of Bmi-1, a transcriptional repressor implicated in the etiology of prostate cancer induced by loss of PTEN function, and the repression of the IGF-IR. The therapeutic implications are significant because combined inhibition of NRP2 and IGF-IR overcomes the resistance induced by targeting each receptor individually. Cancer Discov; 2(10); 906–21. ©2012 AACR.This article is highlighted in the In This Issue feature, p. 857.

PDF file - 2.5MB, Correlation of NRP2 expression in prostate cancer with clinical parameters and expression of VEGFRs

PDF file - 536K, Invasive behavior of cell lines used and assessment of NRP2 expression in prostate glands and cancer

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PDF file - 484K, Evaluation of the contribution of FOXO1 to NRP2 expression

XLS file - 23KB, Supplementary Table S3: Proteins.

PDF file - 2259KB, Supplementary Figure S1. Phenotypic changes consistent with EMT in cE1 culture with CAF CM. A, The greatest observed amount of EMT-like morphology was seen in SCmed cE1 cells grown with CAF CM treatment after 14 days in 3-D Matrigel assays. Representative bright field images of the observed morphologies are shown. B, CAF CM treatment did not lead to increased ability to form spheroids or undergo morphologic changes akin to EMT in SCnone, compared to SCmed. CAFs 1 and 3 were used in independent repeats. Bar 100 μm. Supplementary Figure S2. Ammonium sulfate precipitation of conditioned media proteins. A, Potential mediators of EMT-CSC lineage present in the CAF CM have been enriched using several rounds of ammonium sulfate precipitation followed by in vitro biological assays for protein activity affecting the EMT16 sensitive fraction of the cE1 cell line, SCmed, using the three-dimensional Matrigel culture system. The 40% AS-enriched fraction from the CAF CM had the most extensive EMT-like morphology after 14 days of culture, compared to vehicle control and NPF control. Bright field images were taken at 40x magnification. Bar 100 μm. B, Biological activity for each CAF CM fraction was assayed using the in vitro spheroid formation assay with cE1 SCmed. In agreement with the data in A, SCmed cells treated with CAF CM 40% AS-fraction displayed the highest propensity toward EMT based on quantitative real time PCR analysis of EMT transcription factors (Snail, Slug and Twist), stem cell transcription factors (Oct4, Sox2 and Nanog) and decrease in E-Cadherin. Ratios are expressed as fold change normalized to β-actin expression. CAFs 1 and 3 were used in independent repeats. Supplementary Figure S3. A, Dose response to AnxA1 nAb in the presence of AnxA1 enriched CAF CM AS fraction. CAFs 1 and 3 were used in independent repeats. B, Fpr-rs1 is found in greatest abundance in cE1 cells. ?, P < 0.05; *, P

PDF file - 107KB, Supplementary Table S1. Antibodies and their dilutions for immunohistochemistry (IHC), immunofluorescence (IF), western blot (WB) and cell culture neutralizing antibody assays are listed. Supplementary Table S2. Real Time Quantitative Reverse Transcription PCR Primer List.

PDF file - 582K, Analysis of NRP2, IGF-1R and p-AKT in consecutive sections of prostate cancer specimens

Supplementary Table 1 from Cancer-Associated Fibroblasts Enhance the Gland-Forming Capability of Prostate Cancer Stem Cells

Supplementary Figures 1 and 2 from Bone Morphogenetic Protein 7 Protects Prostate Cancer Cells from Stress-Induced Apoptosis via Both Smad and c-Jun NH2-Terminal Kinase Pathways

Supplementary Figures 1-2 from Inactivation of Apc in the Mouse Prostate Causes Prostate Carcinoma

Osteopontin is a secreted glycosylated phosphoprotein known to be involved in numerous physiologic functions and associated with the late stages of various cancers. We used preneoplastic and neoplastic mouse models of prostate cancer to determine the onset of elevated expression of osteopontin in the development of this disease. Osteopontin alterations occurred early in the disease with dysregulated expression observed in lesions of low-grade prostatic intraepithelial neoplasia (PIN). Over time, osteopontin expressing dysplastic cells seemed to increase in number in high-grade PIN and increased further in adenocarcinoma, and in metastasis, almost all of the cancer cells immunohistochemically stained positive for osteopontin overexpression. We examined the biological properties of human prostate cancer cell lines LNCaP and PC-3, in which osteopontin overexpression was achieved via lentiviral gene transduction. Evidence was obtained that osteopontin could contribute to a proliferative advantage in both cell types, although more significantly in LNCaP than PC-3. Osteopontin also influenced their in vitro invasive ability, and again, most strikingly in the weakly oncogenic LNCaP. Furthermore, excess osteopontin induced the LNCaP cells to acquire a strong intravasation potential in vivo in the chicken embryo chorioallantoic membrane assay for blood vessel penetration. These results establish a correlation between an increased gradient of osteopontin expression throughout the stages of murine prostate cancer, beginning from the preneoplastic lesions to distant metastases that suggests a proliferative and invasive advantages to those prostate tumor cells overexpressing osteopontin. Together, these findings support a strategy designed to target osteopontin in the context of prostate cancer therapy. (Cancer Res 2006; 66(2): 883-8)

Supplementary Table 1, Figures 1-2 from A Novel Bone Morphogenetic Protein Signaling in Heterotypic Cell Interactions in Prostate Cancer

Pathways mediated by p53 and Rb are frequently altered in aggressive human cancers, including prostate carcinoma. To test directly the roles of p53 and Rb in prostate carcinogenesis, we have conditionally inactivated these genes in the prostate epithelium of the mouse. Inactivation of either p53 or Rb leads to prostatic intraepithelial neoplasia developing from the luminal epithelium by 600 days of age. In contrast, inactivation of both genes results in rapidly developing (median survival, 226 days) carcinomas showing both luminal epithelial and neuroendocrine differentiation. The resulting neoplasms are highly metastatic, resistant to androgen depletion from the early stage of development, and marked with multiple gene expression signatures commonly found in human prostate carcinomas. Interestingly, gains at 4qC3 and 4qD2.2 and loss at 14qA2-qD2 have been consistently found by comparative genomic hybridization. These loci contain such human cancer–related genes as Nfib, L-myc, and Nkx3.1, respectively. Our studies show a critical role for p53 and Rb deficiency in prostate carcinogenesis and identify likely secondary genetic alterations. The new genetically defined model should be particularly valuable for providing new molecular insights into the pathogenesis of human prostate cancer. (Cancer Res 2006; 66(16): 7889-98)

Supplementary Figure Legends from Synergy of p53 and Rb Deficiency in a Conditional Mouse Model for Metastatic Prostate Cancer

Signals originating from cancer-associated fibroblasts (CAF) may positively regulate proliferation and tumorigenicity in prostate cancer. In this study, we investigated whether CAFs may regulate the biology of prostate cancer stem cells (CSC) by using a conditional Pten deletion mouse model of prostate adenocarcinoma to isolate both CAF cultures and CSC-enriched cell fractions from the tumors. CSCs that were isolated possessed self-renewal, spheroid-forming, and multipotential differentiation activities in tissue culture, segregating with a cell fraction exhibiting a signature expression phenotype, including SCA-1 (high), CD49f (high), CK5 (high), p63 (high), Survivin (high), RUNX2 (high), CD44 (low), CD133 (low), CK18 (low), and Androgen Receptor (low). CSC spheroid–forming efficiency was differentially influenced by the nature of fibroblasts in a coculture system: Compared with mouse urogenital sinus mesenchyme or normal prostate fibroblasts, CAFs enhanced spheroid formation, with the spheroids displaying generally larger sizes and more complex histology. Graft experiments showed that CSCs admixed with CAFs produced prostatic glandular structures with more numerous lesions, high proliferative index, and tumor-like histopathologies, compared with those formed in the presence of normal prostate fibroblasts. Together, our findings underscore a significant role of CAFs in CSC biology. Cancer Res; 70(18); 7294–303. ©2010 AACR.

We examined the effect of the extracellular bone morphogenetic protein (BMP) 2 and 7, which are up-regulated in the prostate adenocarcinomas of the conditional Pten deletion mouse model, on primary cultures of cancer-associated fibroblasts (CAF) derived from these tumors. In the CAF, we show that BMP2 or BMP7, but not transforming growth factor β-1, can strikingly stimulate secretion of stromal cell–derived factor-1 (SDF-1), also known as CXCL12. The CAF cells express type I and type II BMP receptors as well as the receptor for SDF-1, CXCR4. SDF-1 activation is associated with BMP-induced Smad phosphorylation, and the stimulatory effect is blocked by BMP antagonist, noggin. The findings that BMP treatment can increase SDF-1 pre-mRNA levels in a time-dependent manner and actinomycin D treatment can abolish stimulatory effect of BMP suggest a transcriptional modulation of SDF-1 by BMP signaling. Using a human microvascular endothelial cell line, we show that SDF-1 present in the conditioned medium from the stimulated CAF can significantly induce tube formation, an effect relating to angiogenic function. Furthermore, we found that BMP2 can also protect the CAF from serum starvation–induced apoptosis independent of SDF-1, implying that BMP may induce other factors to sustain the survival of these cells. In short, this report establishes a novel BMP-SDF-1 axis in the prostate tumor along with a new prosurvival effect of BMP that when considered together with our previously described oncogenic properties of BMP indicate a circuitry for heterotypic cell interactions potentially critical in prostate cancer. [Cancer Res 2008;68(1):198–205]

Alterations of the Wnt/β-catenin signaling pathway are positively associated with the development and progression of human cancer, including carcinoma of the prostate. To determine the role of activated Wnt/β-catenin signaling in mouse prostate carcinogenesis, we created a mouse prostate tumor model using probasin-Cre–mediated deletion of Apc. Prostate tumors induced by the deletion of Apc have elevated levels of β-catenin protein and are highly proliferative. Tumor formation is fully penetrant and follows a consistent pattern of progression. Hyperplasia is observed as early as 4.5 weeks of age, and adenocarcinoma is observed by 7 months. Continued tumor growth usually necessitated sacrifice between 12 and 15 months of age. Despite the high proliferation rate, we have not observed metastasis of these tumors to the lymph nodes or other organs. Surgical castration of 6-week-old mice inhibited tumor formation, and castration of mice with more advanced tumors resulted in the partial regression of specific prostate glands. However, significant areas of carcinoma remained 2 months postcastration, suggesting that tumors induced by Apc loss of function are capable of growth under conditions of androgen depletion. We conclude that the prostate-specific deletion of Apc and the increased expression of β-catenin associated with prostate carcinoma suggests a role for β-catenin in prostate cancer and offers an appropriate animal model to investigate the interaction of Wnt signaling with other genetic and epigenetic signals in prostate carcinogenesis. [Cancer Res 2007;67(6):2490–6]

Supplementary Tables S1 & S2 from Synergy of p53 and Rb Deficiency in a Conditional Mouse Model for Metastatic Prostate Cancer

Supplementary Figure Legends 1-2 from Inactivation of Apc in the Mouse Prostate Causes Prostate Carcinoma

Supplementary Figure S1 from Increased Expression of Osteopontin Contributes to the Progression of Prostate Cancer

Supplementary Figure 1 from Cancer-Associated Fibroblasts Enhance the Gland-Forming Capability of Prostate Cancer Stem Cells

We reported earlier that exposure to exogenous bone morphogenetic protein 7 (BMP7) could strongly inhibit serum starvation–induced apoptosis to C4-2B cell line, a variant of the LNCaP human prostate cancer cell line with propensity for bone metastasis. Whereas serum starvation suppressed the expression of survivin, a member of the inhibitor of apoptosis protein family, its expression was sustained in the presence of BMP7. In this study, we present evidence that BMP7 exposure up-regulated survivin promoter activity, an effect that was associated with activation of Smad, and could be repressed by dominant-negative Smad5. Additionally, serum starvation–induced suppression of c-jun NH2-terminal kinase (JNK) activity in C4-2B cells could be mostly restored by BMP7, and a JNK inhibitor could counteract the antiapoptotic effect of BMP7, without a significant effect on the level of survivin expression. Thus, we identified JNK pathway as another signaling mode for the antiapoptotic function of BMP7. To test the effect of endogenous up-regulation of BMP7, we genetically modulated the C4-2B cell line to overexpress BMP7 protein. Not only was this altered cell line resistant to serum starvation–induced apoptosis but it also exhibited patterns of Smad activation, survivin up-regulation, and JNK activation similar to those of the parental C4-2B cells exposed to exogenous BMP7. Consistent with these in vitro findings of BMP7 action, we acquired correlative results of Smad activation, survivin expression, and JNK activation in the progression of prostate cancer in the conditional Pten deletion mouse model, in which we first obtained the evidence of BMP7 overexpression. (Cancer Res 2006; 66(8): 4285-90)

Supplementary Figures S1 & S2 from Synergy of p53 and Rb Deficiency in a Conditional Mouse Model for Metastatic Prostate Cancer

The rate of dilepton emission from a magnetized hot hadronic medium is calculated in the framework of real time formalism of finite temperature field theory. We evaluate the one loop self-energy of neutral rho-meson containing thermo-magnetic propagators for the charged pions in the loop. The in-medium thermo-magnetic spectral function of rho obtained by solving the Dyson-Schwinger equation is shown to be proportional to the dilepton production rate. The study of the analytic structure of the neutral rho-meson spectral function in such a medium shows that in addition to the usual contribution coming from the Unitary cut beyond the two-pion threshold there is a non-trivial yield in the low invariant mass region originating due to the fact that the charged pions occupy different Landau levels before and after scattering with the neutral rho-meson and is purely a finite magnetic field effect., Comment: Version published in Physical Review D

We investigate magnetic field $(eB)$ dependence of constituent quark mass, the longitudinal and transverse pressure as well as the magnetization and magnetic susceptibility of strongly interacting quark matter. We employ the two-flavour Polyakov Nambu--Jona-Lasinio model with the inclusion of the anomalous magnetic moment (AMM) of the quarks at finite temperature $(T)$ and finite quark chemical potential $(\mu_q)$ capturing different stages of chiral phase transition. We find that the transverse pressure, magnetization and magnetic susceptibility become highly oscillatory for large values of $eB$ in the chiral symmetry broken phase. However the oscillations cease to occur at higher values of $T$ and $\mu_q$ when chiral symmetry is (partially) restored and the anisotropic nature of the pressure becomes significant even at smaller values of $eB$. As the inclusion of AMM of the quarks leads to inverse magnetic catalysis of the transition temperature we observe that the variations of transverse pressure, magnetization and magnetic susceptibility are significantly modified in the vicinity of the chiral transition temperature. Furthermore, above the chiral transition temperature the magnetic susceptibility is found to remain positive for a wide range of $eB$ indicating a paramagnetic character of the strongly interacting quark matter. Finally, we have also examined the magnetism of strongly interacting matter in the quarkyonic phase. The obtained results could be useful for a magnetohydrodynamic evolution of hot and dense matter created in heavy-ion collisions., Comment: Version Published in Physical Review D

This paper does a comparative study of the socio-economic condition of the artisanal subsistence marine fishers of the three locations of India namely Sundarbans, Chennai and Paradeep. The small-scale artisanal marine fishers of Chandrapur, Gobindapur, Kalinagar, Ganeshpur villages of Kakdwip Subdivision, Sundarbans along with that of Badakalikhada, Bijaychandrapur, Light House and Kharinali, of Paradeep and Nochi Nagar, Light House, Foreshore Estate, Mandaveli, Mylapore of Chennai were asked the standardised questions. Sundarbans, Paradeep and Chennai have three distinct socio-economic conditions prevailing due to the privatisation of the fishery industry and the distinct difference in income.The purpose of this study is to find the socio-economic condition of the artisanal marine fishers of the Sundarbans, Paradeep and Chennai. A descriptive rapid sampling method has been followed, where the fishers have been questioned about their respective socio-economic status. The paper studies the income difference and the effect of the usages of motorised boats in two three-year slots 2015-2018 and 2018-2020 affluence due to more exposure to the technical changes. The study found that the Chennai fishers were more adept to take the advantage of the new technologies of marine fishing thus far more economical stable, the Paradeep fishers were second in economic stability due to their comparative adaptability. The Sundarbans fishers were least adept with the technological changes in marine fishing, consequently they are least affluent.

We evaluate the dilepton production rate (DPR) from hot and dense chirally asymmetric quark matter. The presence of a finite chiral chemical potential (CCP) in the electromagnetic spectral function results in the appearance of new cut structures signifying additional scattering processes in the medium which leads to a significant enhancement in the DPR at lower values of invariant mass. The constituent quark mass evaluated using a 3-flavour Nambu--Jona-Lasinio model is also non-trivially affected by the CCP. These are found to result in a continuous dilepton production rate as a function of the invariant mass for higher values of temperature and baryonic chemical potential., Comment: Version published in Physical Review D

We study the production of heavy-flavor decay muons (HFM) in proton–proton ([Formula: see text]–[Formula: see text]) and heavy-ion ([Formula: see text]–[Formula: see text]) collisions using Angantyr model in PYTHIA8 at Large Hadron Collider (LHC) energies. These studies have been performed for three colliding systems: small ([Formula: see text]–[Formula: see text]), intermediate (Xe–Xe) and large (Pb–Pb). The works have been performed in [Formula: see text]–[Formula: see text] (with [Formula: see text] = 2.76, 5.02, 7.0 and 13[Formula: see text]TeV), Xe–Xe (with [Formula: see text][Formula: see text]=[Formula: see text]5.44[Formula: see text]TeV) and Pb–Pb (with [Formula: see text][Formula: see text]=[Formula: see text]2.76 and 5.02[Formula: see text]TeV) collision systems. The production of heavy-flavor (charm and bottom quarks) decay muons in [Formula: see text]–[Formula: see text] and [Formula: see text]–[Formula: see text] collisions is measured in the forward rapidity [Formula: see text]. The standard statistical uncertainty is computed for each case as [Formula: see text]. In the study of [Formula: see text]-distribution of HFM at forward rapidity ([Formula: see text]) using Angantyr model, we find that simulation results agree with ALICE data reasonably well for [Formula: see text]–[Formula: see text] and [Formula: see text]–[Formula: see text] collisions. The comparison of simulation results with ALICE data for [Formula: see text]-distribution at [Formula: see text] and 7.0[Formula: see text]TeV in [Formula: see text]–[Formula: see text] collision has also been illustrated. In addition to the particle spectra, a more quantitative model-to-data comparison can be obtained through the observable [Formula: see text]. We, thus, calculate [Formula: see text] of HFM in this work for [Formula: see text] and 5.02[Formula: see text]TeV. The simulated [Formula: see text] results overestimate the ALICE data. [Formula: see text] estimation as a function of [Formula: see text] shows almost constant values and close to unity at lower energy ([Formula: see text][Formula: see text]=[Formula: see text]2.76[Formula: see text]TeV), but substantially lower than unity at comparatively higher energy ([Formula: see text][Formula: see text]=[Formula: see text]5.02[Formula: see text]TeV). We also predict for the HFM [Formula: see text] spectrum and [Formula: see text]-distribution in O–O collision at [Formula: see text][Formula: see text]=[Formula: see text]6.37[Formula: see text]TeV for 0–10[Formula: see text] collision centrality. These simulated spectra are illustrated for ALICE Run3 experiment and they comply with other HFM ALICE measurements.

The photon self-energy from chirally imbalanced quark matter is evaluated at finite temperature and density using the real time formulation of thermal field theory. The analytic structure is explored in detail exposing the cut structure which corresponds to a variety of physical scattering and decay processes in the medium and their thresholds. The mass of the quarks in the chiral symmetry broken phase are obtained from the gap equation of the Nambu--Jona-Lasinio model. It is found that, in presence of finite chiral chemical potential, the chiral condensate tends to get stronger at low temperature while the opposite is observed at high values of temperature. A continuous spectrum is obtained for the electromagnetic spectral function and this is purely a finite chiral chemical potential effect., Comment: Version Published in Physical Review D

We incorporate the anomalous magnetic moment (AMM) of quarks in the framework of PNJL model to study hot and dense magnetised matter with chiral imbalance. For this purpose, the eigen energy solution of the Dirac equation is obtained in presence of constant background magnetic field and chiral chemical potential (CCP) along with the minimal anomalous magnetic moment interaction of the fermion. Although there is a marginal enhancement in the IMC behaviour of the quark condensate due to the combined effects of AMM and CCP, we find that the overall behaviour of the Polyakov loop and the chiral charge density is dominated by the chiral chemical potential. It is further shown that the AMM effects in presence of CCP remains insignificant even after consideration of thermo-magnetically modified moments., Comment: Version published in European Physical Journal A