Probing the luminal microenvironment of reconstituted epithelial microtissues.

Polymeric microparticles can serve as carriers or sensors to instruct or characterize tissue biology. However, incorporating microparticles into tissues for in vitro assays remains a challenge. We exploit three-dimensional cell-patterning technologies and directed epithelial self-organization to deliver microparticles to the lumen of reconstituted human intestinal microtissues. We also develop a novel pH-sensitive microsensor that can measure the luminal pH of reconstituted epithelial microtissues. These studies offer a novel approach for investigating luminal microenvironments and drug-delivery across epithelial barriers

Laser powder bed fusion additive manufacturing of oxide dispersion strengthened steel using gas atomized reaction synthesis powder

Mechanically alloyed Fe-based alloys with oxide dispersion strengthening have largely dropped out of the marketplace due to high cost related to problems with complex and unreliable processing. Nevertheless, the desirable properties of oxide dispersion strengthened (ODS) steels have motivated research on alternate processing routes aimed at improving processing simplicity and reliability. Powders produced by gas atomization reaction synthesis (GARS) consist of stable Fe-Y intermetallic phases and a Cr surface oxide layer that acts as a chemical reservoir during solid-state processing and heat treatment to form a high density of nano-scale oxides. This research explores the use of Fe GARS powders, with 15 wt% Cr with micro-alloyed additions of 0.15 wt% Y and 0.10% Ti, in laser powder bed fusion (LPBF) additive manufacturing (AM), and evaluates the effectiveness of oxide dispersoid formation in the liquid melt pool. Additional oxygen was introduced by varying the LPBF chamber atmospheres using Ar, Ar + 1 wt% O, Ar + 5 wt% O, and air. Characterization of LPBF consolidated solids demonstrated the formation of a high density of nano-scale Y-Ti oxides in the build microstructures from the GARS precursor powders.This is a manuscript of an article published as Horn, Timothy, Christopher Rock, Djamel Kaoumi, Iver Anderson, Emma White, Tim Prost, Joel Rieken et al. "Laser powder bed fusion additive manufacturing of oxide dispersion strengthened steel using gas atomized reaction synthesis powder." Materials & Design 216 (2022): 110574. DOI: 10.1016/j.matdes.2022.110574. Copyright 2022 The Authors. Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0). Posted with permission. DOE Contract Number(s): AC02-07CH11358; AR0001379; AC02-06CH11357; AC05-76RL01830; ECCS-2025064

Summary Paper on Underactive Bladder from the European Association of Urology Guidelines on Non-neurogenic Male Lower Urinary Tract Symptoms

International audienc

PHIP as a therapeutic target for driver-negative subtypes of melanoma, breast, and lung cancer

The identification and targeting of key molecular drivers of melanoma and breast and lung cancer have substantially improved their therapy. However, subtypes of each of these three common, lethal solid tumors lack identified molecular drivers, and are thus not amenable to targeted therapies. Here we show that pleckstrin homology domain-interacting protein (PHIP) promotes the progression of these "driver-negative" tumors. Suppression of PHIP expression significantly inhibited both tumor cell proliferation and invasion, coordinately suppressing phosphorylated AKT, cyclin D1, and talin1 expression in all three tumor types. Furthermore, PHIP's targetable bromodomain is functional, as it specifically binds the histone modification H4K91ac. Analysis of TCGA profiling efforts revealed PHIP overexpression in triple-negative and basal-like breast cancer, as well as in the bronchioid subtype of nonsmall cell lung cancer. These results identify a role for PHIP in the progression of melanoma and breast and lung cancer subtypes lacking identified targeted therapies. The use of selective, anti-PHIP bromodomain inhibitors may thus yield a broad-based, molecularly targeted therapy against currently nontargetable tumors

PHIP as a therapeutic target for driver-negative subtypes of melanoma, breast, and lung cancer

The identification and targeting of key molecular drivers of melanoma and breast and lung cancer have substantially improved their therapy. However, subtypes of each of these three common, lethal solid tumors lack identified molecular drivers, and are thus not amenable to targeted therapies. Here we show that pleckstrin homology domain-interacting protein (PHIP) promotes the progression of these "driver-negative" tumors. Suppression of PHIP expression significantly inhibited both tumor cell proliferation and invasion, coordinately suppressing phosphorylated AKT, cyclin D1, and talin1 expression in all three tumor types. Furthermore, PHIP's targetable bromodomain is functional, as it specifically binds the histone modification H4K91ac. Analysis of TCGA profiling efforts revealed PHIP overexpression in triple-negative and basal-like breast cancer, as well as in the bronchioid subtype of nonsmall cell lung cancer. These results identify a role for PHIP in the progression of melanoma and breast and lung cancer subtypes lacking identified targeted therapies. The use of selective, anti-PHIP bromodomain inhibitors may thus yield a broad-based, molecularly targeted therapy against currently nontargetable tumors

PHIP drives glioblastoma motility and invasion by regulating the focal adhesion complex

The invasive behavior of glioblastoma is essential to its aggressive potential. Here, we show that pleckstrin homology domain interacting protein (PHIP), acting through effects on the force transduction layer of the focal adhesion complex, drives glioblastoma motility and invasion. Immunofluorescence analysis localized PHIP to the leading edge of glioblastoma cells, together with several focal adhesion proteins: vinculin (VCL), talin 1 (TLN1), integrin beta 1 (ITGB1), as well as phosphorylated forms of paxillin (pPXN) and focal adhesion kinase (pFAK). Confocal microscopy specifically localized PHIP to the force transduction layer, together with TLN1 and VCL. Immunoprecipitation revealed a physical interaction between PHIP and VCL. Targeted suppression of PHIP resulted in significant down-regulation of these focal adhesion proteins, along with zyxin (ZYX), and produced profoundly disorganized stress fibers. Live-cell imaging of glioblastoma cells overexpressing a ZYX-GFP construct demonstrated a role for PHIP in regulating focal adhesion dynamics. PHIP silencing significantly suppressed the migratory and invasive capacity of glioblastoma cells, partially restored following TLN1 or ZYX cDNA overexpression. PHIP knockdown produced substantial suppression of tumor growth upon intracranial implantation, as well as significantly reduced microvessel density and secreted VEGF levels. PHIP copy number was elevated in the classical glioblastoma subtype and correlated with elevated EGFR levels. These results demonstrate PHIP's role in regulating the actin cytoskeleton, focal adhesion dynamics, and tumor cell motility, and identify PHIP as a key driver of glioblastoma migration and invasion

Insulin-like Growth Factor Messenger RNA-binding Protein 3 Expression Helps Prognostication in Patients with Upper Tract Urothelial Carcinoma

International audienceBACKGROUND: Upper tract urothelial carcinoma (UTUC) is a clinically heterogeneous disease that lacks high-quality trials that provide definitive prognostic markers. Insulin-like growth factor messenger RNA binding protein 3 (IMP3) has been associated with outcomes in urothelial carcinoma of the bladder but was not yet studied in UTUC. OBJECTIVE: To evaluate the association of the oncofetal protein IMP3 with oncologic outcomes in patients with UTUC treated with radical nephroureterectomy (RNU). DESIGN, SETTING, AND PARTICIPANTS: We investigated the expression of IMP3 and its association with clinical outcomes using tissue microarrays constructed from 622 patients treated with RNU at seven international institutions between 1991 and 2008. INTERVENTION: All patients were diagnosed with UTUC and underwent RNU. OUTCOME MEASUREMENT AND STATISTICAL ANALYSIS: Uni- and multivariable Cox regression analyses evaluated the association of IMP3 protein expression with disease recurrence, cancer-specific mortality, and all-cause mortality. RESULTS AND LIMITATIONS: IMP3 was expressed in 12.2% of patients with UTUC (n=76). The expression was tumor specific and correlated with higher stages/grades. Within a median follow-up of 27 mo (interquartile range [IQR]: 12-53), 191 patients (25.4%) experienced disease recurrence, and 165 (21.9%) died of the disease. Patients with IMP3 demonstrated significantly worse recurrence-free survival (27.4% vs 75.1%; p\textless0.01), cancer-specific survival (34.5% vs 78.9%; p\textless0.01), and overall survival (15.6% vs 64.8%; p\textless0.01) at 5 yr compared with those without IMP3. In multivariable Cox regression analyses, which adjusted for the effects of standard clinicopathologic features, IMP3expression was independently associated with disease recurrence (hazard ratio [HR]: 1.87; p\textless0.01), cancer-specific mortality (HR: 2.15; p\textless0.01), and all-cause mortality (HR: 2.07; p\textless0.01). Major limitations include the retrospective design and relatively short follow-up time. CONCLUSIONS: IMP3 expression is independently associated with disease recurrence, cancer-specific mortality, and all-cause mortality in UTUC. IMP3 may help improve risk stratification and prognostication of UTUC patients treated with RNU