FAP-overexpressing fibroblasts produce an extracellular matrix that enhances invasive velocity and directionality of pancreatic cancer cells

<p>Abstract</p> <p>Background</p> <p>Alterations towards a permissive stromal microenvironment provide important cues for tumor growth, invasion, and metastasis. In this study, Fibroblast activation protein (FAP), a serine protease selectively produced by tumor-associated fibroblasts in over 90% of epithelial tumors, was used as a platform for studying tumor-stromal interactions.</p> <p>We tested the hypothesis that FAP enzymatic activity locally modifies stromal ECM (extracellular matrix) components thus facilitating the formation of a permissive microenvironment promoting tumor invasion in human pancreatic cancer.</p> <p>Methods</p> <p>We generated a tetracycline-inducible FAP overexpressing fibroblastic cell line to synthesize an <it>in vivo</it>-like 3-dimensional (3D) matrix system which was utilized as a stromal landscape for studying matrix-induced cancer cell behaviors. A FAP-dependent topographical and compositional alteration of the ECM was characterized by measuring the relative orientation angles of fibronectin fibers and by Western blot analyses. The role of FAP in the matrix-induced permissive tumor behavior was assessed in Panc-1 cells in assorted matrices by time-lapse acquisition assays. Also, FAP⁺matrix-induced regulatory molecules in cancer cells were determined by Western blot analyses.</p> <p>Results</p> <p>We observed that FAP remodels the ECM through modulating protein levels, as well as through increasing levels of fibronectin and collagen fiber organization. FAP-dependent architectural/compositional alterations of the ECM promote tumor invasion along characteristic parallel fiber orientations, as demonstrated by enhanced directionality and velocity of pancreatic cancer cells on FAP⁺matrices. This phenotype can be reversed by inhibition of FAP enzymatic activity during matrix production resulting in the disorganization of the ECM and impeded tumor invasion. We also report that the FAP⁺ matrix-induced tumor invasion phenotype is β₁-integrin/FAK mediated.</p> <p>Conclusion</p> <p>Cancer cell invasiveness can be affected by alterations in the tumor microenvironment. Disruption of FAP activity and β₁-integrins may abrogate the invasive capabilities of pancreatic and other tumors by disrupting the FAP-directed organization of stromal ECM and blocking β₁-integrin dependent cell-matrix interactions. This provides a novel preclinical rationale for therapeutics aimed at interfering with the architectural organization of tumor-associated ECM. Better understanding of the stromal influences that fuel progressive tumorigenic behaviors may allow the effective future use of targeted therapeutics aimed at disrupting specific tumor-stromal interactions.</p

Cdc42 Regulates Extracellular Matrix Remodeling in Three Dimensions*

Extracellular matrix (ECM) actively participates in normal cell regulation and in the process of tumor progression. The Rho GTPase Cdc42 has been shown to regulate cell-ECM interaction in conventional two-dimensional culture conditions by using dominant mutants of Cdc42 in immortalized cell lines that may introduce nonspecific effects. Here, we employ three-dimensional culture systems for conditional gene targeted primary mouse embryonic fibroblasts that better simulate the reciprocal and adaptive interactions between cells and surrounding matrix to define the role of Cdc42 signaling pathways in ECM organization. Cdc42 deficiency leads to a defect in global cell-matrix interactions reflected by a decrease in collagen gel contraction. The defect is associated with an altered cell-matrix interaction that is evident by morphologic changes and reduced focal adhesion complex formation. The matrix defect is also associated with a reduction in synthesis and activation of matrix metalloproteinase 9 (MMP9) and altered fibronectin deposition patterning. A Cdc42 mutant rescue experiment found that downstream of Cdc42, p21-activated kinase (PAK), but not Par6 or WASP, may be involved in regulating collagen gel contraction and fibronectin organization. Thus, in addition to the previously implicated roles in intracellular regulation of actin organization, proliferation, and vesicle trafficking, Cdc42 is essential in ECM remodeling in three dimensions

Fixed Drug Eruption Due to Allopurinol: Positive Oral Provocation

A fixed drug eruption (FDE) is characterized by the presence of a solitary or multiple, pruritic, well-circumscribed, erythematous plaques. These lesions have tendency to recur at same sites and heal with residual hyperpigmenation. With repeated attacks, the size and/or number of the lesions may increase. So far, more than 100 drugs have been implicated in causing FDEs, including ibuprofen, sulfonamide, naproxen, and tetracylines. FDE caused by allopurinol has been rarely reported in the literature, but there has been no confirmed case based on oral provocation test. Herein, we report a case of FDE in which the lesions recurred whenever allopurinol was administered for the treatment of gout. A 64-year-old male experienced repeated episodes of well-demarcated dusky erythematous patches on the whole body for 2 months. He took allopurinol intermittently for amelioration of his gout symptom, but denied other medication history. Pruritic erythematous edema developed on the previous lesions 12 hours after oral provocation of 200 mg of allopurinol

Fixed Drug Eruption due to Iopromide (Ultravist®)

A 69-year-old male presented with several painful erythematous patches on both palms and trunk several days after receiving iopromide (Ultravist®, Shering, Berlin, Germany). A fixed drug eruption (FDE) due to iopromide was suspected clinically. However, at that time, the patch test with iopromide at the lesion site gave negative results. Three years later, the patient was mistakenly administered iopromide again and patches with vesicles recurred on the same sites as well as on the genitalia. This episode was repeated once again after 1 year. In all episodes, the skin lesion resolved after application of topical steroids. Although a patch test with iopromide was negative in our case, we made a diagnosis of FDE due to iopromide because the skin lesions occurred again at the previously involved area after re-exposure to iopromide. To date, only three cases of FDE caused by non-ionic monomers have been documented in the English literature. Herein, we report on an interesting case of FDE caused by iopromide

Photodynamic Therapy Combined with CO2 Laser Vaporization on Disseminated Superficial Actinic Porokeratosis: A Report of 2 Cases on the Face

Disseminated superficial actinic porokeratosis (DSAP) is a skin condition that usually shows a poor response to different modalities of treatment. Herein we describe 2 patients with DSAP on the face, each treated with 3 to 4 sessions of photodynamic therapy combined with laser vaporization

Sebaceous Hyperplasia of the Scrotum and Penile Shaft

Sebaceous hyperplasia (SH) is a proliferative abnormality of sebaceous glands, most often seen in elderly individuals. It consists of single or multiple, asymptomatic, small yellow papules with a central depression. The face is most commonly affected, but there have been reports of SH on the chest or genitalia. A lobular array of well-differentiated mature sebaceous lobules is noted on histological preparations. In this report, we describe a patient with SH on the penile shaft and parts of the scrotum

Delayed Migration of Tapered Open-Cell Design Carotid Stent: A Case Report

We present a case of delayed migration of an open-cell design carotid stent, which is a rare complication following carotid artery stenting (CAS). A 65-year-old patient with carotid artery stenosis underwent CAS with an open-cell stent, initially achieving successful deployment. However, 4 months later, the stent migrated and resulted in restenosis. The patient underwent balloon angioplasty and received an additional stent, leading to improved blood flow. The rarity of stent migration, particularly in the absence of risk factors, highlights the need for clinicians to be vigilant and consider early imaging follow-up for patients at risk of this complication after CAS