Proteomic characterisation of prostate cancer intercellular communication reveals cell type-selective signalling and TMSB4X-dependent fibroblast reprogramming

Background: In prostate cancer, the tumour microenvironment (TME) represents an important regulator of disease progression and response to treatment. In the TME, cancer-associated fbroblasts (CAFs) play a key role in tumour progression, however the mechanisms underpinning fbroblast-cancer cell interactions are incompletely resolved. Here, we address this by applying cell type-specifc labelling with amino acid precursors (CTAP) and mass spectrometry (MS)-based (phospho) proteomics to prostate cancer for the frst time. Methods: Reciprocal interactions between PC3 prostate cancer cells co-cultured with WPMY-1 prostatic fbroblasts were characterised using CTAP-MS. Signalling network changes were determined using Metascape and Enrichr and visualised using Cytoscape. Thymosin β4 (TMSB4X) overexpression was achieved via retroviral transduction and assayed by ELISA. Cell motility was determined using Transwell and random cell migration assays and expression of CAF markers by indirect immunofuorescence. Results: WPMY-1 cells co-cultured with PC3s demonstrated a CAF-like phenotype, characterised by enhanced PDGFRB expression and alterations in signalling pathways regulating epithelial-mesenchymal transition, cytoskeletal organisation and cell polarisation. In contrast, co-cultured PC3 cells exhibited more modest network changes, with alterations in mTORC1 signalling and regulation of the actin cytoskeleton. The expression of the actin binding protein TMSB4X was signifcantly decreased in co-cultured WPMY-1 fbroblasts, and overexpression of TMSB4X in fbroblasts decreased migration of cocultured PC3 cells, reduced fbroblast motility, and protected the fbroblasts from being educated to a CAF-like phenotype by prostate cancer cells. Conclusions: This study highlights the potential of CTAP-MS to characterise intercellular communication within the prostate TME and identify regulators of cellular crosstalk such as TMSB4X.Yunjian Wu, Kimberley C. Clark, Elizabeth V. Nguyen, Birunthi Niranjan, Lisa G. Horvath, Renea A. Taylor, Roger J. Dal

A Small Molecule that Induces Intrinsic Pathway Apoptosis with Unparalleled Speed

Apoptosis is generally believed to be a process thatrequires several hours, in contrast to non-programmed forms of cell death that can occur in minutes. Our findings challenge the time-consuming nature of apoptosis as we describe the discovery and characterization of a small molecule, named Raptinal, which initiates intrinsic pathway caspase-dependent apoptosis within minutes in multiple cell lines. Comparison to a mechanistically diverse panel of apoptotic stimuli reveals that Raptinal-induced apoptosis proceeds with unparalleled speed. The rapid phenotype enabled identification of the criticalroles of mitochondrial voltage-dependent anion channel function, mitochondrial membrane potential/coupled respiration, and mitochondrial complex I, III, and IV function for apoptosis induction. Use of Raptinal in whole organisms demonstrates its utility for studying apoptosis invivo for a variety of applications. Overall, rapid inducers of apoptosis are powerful tools that will be used in a variety of settings to generate further insight into the apoptotic machinery. Palchaudhuri etal. describe the discovery of a small molecule called "Raptinal" that induces unusually rapid apoptotic cell death via the intrinsic pathway. Their work describes the utility of Raptinal as a tool for apoptosis induction relative to other available small molecules

Branching Fractions for D0 -> K+K- and D0 -> pi+pi-, and a Search for CP Violation in D0 Decays

Using the large hadroproduced charm sample collected in experiment E791 at Fermilab, we have measured ratios of branching fractions for the two-body singly-Cabibbo-suppressed charged decays of the D0: (D0 -> KK)/(D0 -> Kpi) = 0.109 +- 0.003 +- 0.003, (D0 -> pipi)/(D0 -> Kpi) = 0.040 +- 0.002 +- 0.003, and (D0 -> KK)/(D0 -> pipi) = 2.75 +- 0.15 +- 0.16. We have looked for differences in the decay rates of D0 and D0bar to the CP eigenstates K+K- and pi+pi-, and have measured the CP asymmetry parameters A_CP(K+K-) = -0.010 +- 0.049 +- 0.012 and A_CP(pi+pi-) = -0.049 +- 0.078 +- 0.030, both consistent with zero.Comment: 10 Postscript pages, including 2 figures. Submitted to Phys. Lett.

Search for CP Violation in Charged D Meson Decays

We report results of a search for CP violation in the singly Cabibbo-suppressed decays D+ -> K- K+ pi+, phi pi+, K*(892)0 K+, and pi- pi+ pi+ based on data from the charm hadroproduction experiment E791 at Fermilab. We search for a difference in the D+ and D- decay rates for each of the final states. No evidence for a difference is seen. The decay rate asymmetry parameters A(CP), defined as the difference in the D+ and D- decay rates divided by the sum of the decay rates, are measured to be: A(CP)(K K pi) = -0.014 +/- 0.029, A(CP)(phi pi) = -0.028 +/- 0.036, A(CP)(K*(892) K) = -0.010 +/- 0.050, and A(CP)(pi pi pi) = -0.017 +/- 0.042.Comment: 13 pages, 5 figures, 1 table; Elsevier LaTe

Asymmetries between the production of D+ and D- mesons from 500 GeV/c pi- nucleon interactions as a function of xF and pt**2

We present asymmetries between the production of D+ and D- mesons in Fermilab experiment E791 as a function of xF and pt**2. The data used here consist of 74,000 fully-reconstructed charmed mesons produced by a 500 GeV/c pi- beam on C and Pt foils. The measurements are compared to results of models which predict differences between the production of heavy-quark mesons that have a light quark in common with the beam (leading particles) and those that do not (non-leading particles). While the default models do not agree with our data, we can reach agreement with one of them, PYTHIA, by making a limited number of changes to parameters used

Search for Rare and Forbidden Dilepton Decays of the D+, Ds, and D0 Charmed Mesons

We report the results of a search for flavor-changing neutral current, lepton-flavor violating, and lepton-number violating decays of D+, Ds, and D0 mesons (and their antiparticles) into modes containing muons and electrons. Using data from Fermilab charm hadroproduction experiment E791, we examine the pi,l,l and K,l,l decay modes of D+ and Ds and the l+l- decay modes of D0. No evidence for any of these decays is found. Therefore, we present branching-fraction upper limits at 90% confidence level for the 24 decay modes examined. Eight of these modes have no previously reported limits, and fourteen are reported with significant improvements over previously published results.Comment: 12 pages, 3 figures, LaTeX, elsart.cls, epsf.sty, amsmath.sty Submitted to Physics Letters

Measurement of the form-factor ratios for D+ --> K* l nu

The form factor ratios rv=V(0)/A1(0), r2=A2(0)/A1(0) and r3=A3(0)/A1(0) in the decay D+ --> K* l nu, K* -->K-pi+ have been measured using data from charm hadroproduction experiment E791 at Fermilab. From 3034 (595) signal (background) events in the muon channel, we obtain rv=1.84+-0.11+-0.09, r2=0.75+-0.08+-0.09 and, as a first measurement of r3, we find 0.04+-0.33 +-0.29. The values of the form factor ratios rv and r2 measured for the muon channel are combined with the values of rv and r2 that we have measured in the electron channel. The combined E791 results for the muon and electron channels are rv=1.87+-0.08+-0.07 and r2=0.73+-0.06+-0.08.Comment: 9 pages + 3 figures ; submitted to PL

Differential cross sections, charge production asymmetry, and spin-density matrix elements for D*(2010) produced in 500 GeV/c pi^- nucleon interactions

We report differential cross sections for the production of D*(2010) produced in 500 GeV/c pi^- nucleon interactions from experiment E791 at Fermilab, as functions of Feynman-x (x_F) and transverse momentum squared (p_T^2). We also report the D* +/- charge asymmetry and spin-density matrix elements as functions of these variables. Investigation of the spin-density matrix elements shows no evidence of polarization. The average values of the spin alignment are \eta= 0.01 +- 0.02 and -0.01 +- 0.02 for leading and non-leading particles, respectively.Comment: LaTeX2e (elsart.cls). 13 pages, 6 figures (eps files). Submitted to Physics Letters

MET-receptor targeted fluorescent imaging and spectroscopy to detect multifocal papillary thyroid cancer

Purpose: Multifocal disease in PTC is associated with an increased recurrence rate. Multifocal disease (MD) is underdiagnosed with the current gold standard of pre-operative ultrasound staging. Here, we evaluate the use of EMI-137 targeted molecular fluorescence-guided imaging (MFGI) and spectroscopy as a tool for the intra-operative detection of uni- and multifocal papillary thyroid cancer (PTC) aiming to improve disease staging and treatment selection. Methods: A phase-1 study (NCT03470259) with EMI-137 was conducted to evaluate the possibility of detecting PTC using MFGI and quantitative fiber-optic spectroscopy. Results: Fourteen patients underwent hemi- or total thyroidectomy (TTX) after administration of 0.09 mg/kg (n = 1), 0.13 mg/kg (n = 8), or 0.18 mg/kg (n = 5) EMI-137. Both MFGI and spectroscopy could differentiate PTC from healthy thyroid tissue after administration of EMI-137, which binds selectively to MET in PTC. 0.13 mg/kg was the lowest dosage EMI-137 that allowed for differentiation between PTC and healthy thyroid tissue. The smallest PTC focus detected by MFGI was 1.4 mm. MFGI restaged 80% of patients from unifocal to multifocal PTC compared to ultrasound. Conclusion: EMI-137-guided MFGI and spectroscopy can be used to detect multifocal PTC. This may improve disease staging and treatment selection between hemi- and total thyroidectomy by better differentiation between unifocal and multifocal disease. Trial registration: NCT03470259.</p

Mass Splitting and Production of Σc0\Sigma_c^0 and Σc++\Sigma_c^{++} Measured in 500GeV500 {GeV} π−−\pi^- -N Interactions

From a sample of $2722 \pm 78$ $\Lambda_c^+$ decaying to the $pK^-\pi^+$ final state, we have observed, in the hadroproduction experiment E791 at Fermilab, $143 \pm 20$ $\Sigma_c^0$ and $122 \pm 18$ $\Sigma_c^{++}$ through their decays to $\Lambda_c^+ \pi^{\pm}$. The mass difference $M(\Sigma_c^0) - M(\Lambda_c^+$) is measured to be $(167.38\pm 0.29\pm 0.15) {MeV}$; for $M(\Sigma_c^{++}) - M(\Lambda_c^+)$, we find $(167.76\pm 0.29\pm0.15) {MeV}$. The rate of $\Lambda_c^+$ production from decays of the $\Sigma_c$ triplet is (22\pm 2\pm 3) {%} of the total $\Lambda_c^+$ production assuming equal rate of production from all three, as measured for $\Sigma_c^0$ and $\Sigma_c^{++}$. We do not observe a statistically significant $\Sigma_c$ baryon-antibaryon production asymmetry. The $x_F$ and $p_t^2$ spectra of $\Lambda_c^+$ from $\Sigma_c$ decays are observed to be similar to those for all $\Lambda_c^+$'s produced.Comment: 15 pages, uuencoded postscript 3 figures uuencoded, tar-compressed fil