Magaly Martinez-Ferrer

Dermal Transforming Growth Factor-β Responsiveness Mediates Wound Contraction and Epithelial Closure

Stromal-epithelial interactions are important during wound healing. Transforming growth factor-beta (TGF-beta) signaling at the wound site has been implicated in re-epithelization, inflammatory infiltration, wound contraction, and extracellular matrix deposition and remodeling. Ultimately, TGF-beta is central to dermal scarring. Because scarless embryonic wounds are associated with the lack of dermal TGF-beta signaling, we studied the role of TGF-beta signaling specifically in dermal fibroblasts through the development of a novel, inducible, conditional, and fibroblastic TGF-beta type II receptor knockout (Tgfbr2(dermalKO)) mouse model. Full thickness excisional wounds were studied in control and Tgfbr2(dermalKO) back skin. The Tgfbr2(dermalKO) wounds had accelerated re-epithelization and closure compared with controls, resurfacing within 4 days of healing. The loss of TGF-beta signaling in the dermis resulted in reduced collagen deposition and remodeling associated with a reduced extent of wound contraction and elevated macrophage infiltration. Tgfbr2(dermalKO) and control skin had similar numbers of myofibroblastic cells, suggesting that myofibroblastic differentiation was not responsible for reduced wound contraction. However, several mediators of cell-matrix interaction were reduced in the Tgfbr2(dermalKO) fibroblasts, including alpha1, alpha2, and beta1 integrins, and collagen gel contraction was diminished. There were associated deficiencies in actin cytoskeletal organization of vasodilator-stimulated phosphoprotein-containing lamellipodia. This study indicated that paracrine and autocrine TGF-beta dermal signaling mechanisms mediate macrophage recruitment, re-epithelization, and wound contraction.

Chemokine markers predict biochemical recurrence of prostate cancer following prostatectomy

Purpose: Stratifying patients who have a high risk of prostate cancer recurrence following prostatectomy can potentiate the use of adjuvant therapy at an early stage. Inflammation has emerged as a mediator of prostate cancer metastatic progression. We hypothesized that chemokines can be biomarkers for distinguishing patients with high risk for biochemical recurrence of prostate cancer. Experimental Design: In a nested case-control study, 82 subjects developed biochemical recurrence within 5 years of prostatectomy. Prostate tissues from 98 age-matched subjects who were recurrence-free following prostatectomy in the same period were the controls. A high-throughput lectin-based enrichment of prostate tissue enabled multiplex ELISA to identify the expression of three chemokines to discriminate the two patient populations. Results: The expression of CX3CL1 and IL-15 in prostate tissue was associated with 5-year biochemical recurrence-free survival following prostatectomy. However, the expression of chemokine ligand 4 (CCL4) was associated with biochemical recurrence. Multivariable logistic regression model combining preoperative prostate-specific antigen, Gleason score, surgical margin, and seminal vesicle status with the three chemokines doubled the specificity of prediction at 90% sensitivity compared with use of the clinicopathologic variables alone (P < 0.0001). Survival analysis yielded a nomogram that supported the use of CX3CL1, IL-15, and CCL4 in predicting 1-, 3-, and 5-year recurrence-free survival after prostatectomy. Conclusions: Each of the three chemokines can serve as independent predictors of biochemical recurrence. However, the combination of chemokine biomarkers plus clinicopathologic variables discriminated prostatectomy subjects for the probability of biochemical recurrence significantly better than clinicopathologic variables alone.

Nicotinic Signaling Ameliorates Acute Bladder Inflammation Induced by Protamine Sulfate or Cyclophosphamide

PURPOSE Nicotinic afferent pathways may be involved in the regulation of bladder inflammation. Based on that hypothesis we investigated the role of nicotinic signaling in a comparative analysis of 2 models of experimental bladder inflammation using protamine sulfate and cyclophosphamide. MATERIALS AND METHODS Protamine sulfate and cyclophosphamide were used to induce acute bladder inflammation. Nicotinic agonists and antagonists were given concomitant to the bladder inflammatory agents. Changes in bladder inflammation were measured histologically by a pathologist and through the expression of inflammatory genes. RESULTS Histologically cyclophosphamide induced more inflammatory changes than protamine sulfate during acute bladder inflammation. Antagonizing nicotinic signaling with mecamylamine induced further inflammatory changes on histology when used with cyclophosphamide but not with protamine sulfate. However, antagonizing nicotinic signaling in combination with protamine sulfate induced greater increases in mRNA expression of the inflammatory cytokine interleukin-6 compared to cyclophosphamide and mecamylamine combination treatments. The activation of nicotinic signaling attenuated acute bladder inflammation by protamine sulfate and cyclophosphamide independently through the down-regulation of increased interleukin-6 expression. CONCLUSIONS Acutely cyclophosphamide treatment results in a greater frank bladder inflammation model in mice than protamine sulfate. However, cholinergic signaling can inhibit inflammation by either mechanism of induced bladder injury. Interleukin-6 gene expression is present and it can be regulated by afferent neuronal signaling even in the absence of observed histological changes in acute bladder inflammatory models.

IL-15 regulates migration, invasion, angiogenesis and genes associated with lipid metabolism and inflammation in prostate cancer

Prostate cancer (PCa) is the most commonly diagnosed non-cutaneous cancer. In the United States it is second leading cause of cancer related deaths in men. PCa is often treated via radical prostatectomy (RP). However, 15–30% of the patients develop biochemical recurrence (i.e. increased serum prostate specific antigen (PSA) levels). Interleukin-15 (IL-15) is a secreted cytokine found over expressed in patients with recurrence-free survival after RP. In our study, we aim to determine the role of IL-15 in PCa using in vitro and in vivo models, and gene expression analysis. PC3 (androgen-independent) and 22RV1 (androgen-dependent) cell lines were treated with IL-15 at 0.0013 ng/mL and 0.1 ng/mL. Tumor growth was evaluated using an orthotopic xenograft model. The anterior prostate lobes of SCID mice were injected with 250,000 22RV1 cells and IL-15 was administered bi-weekly with intraperitoneal (IP) injections during 4 weeks. Tumor tissue was collected for immunohistochemical and gene expression analysis. To study changes in gene expression, we looked at “Tumor Metastasis” and “PI3K pathway” using commercially available PCR arrays. In addition, we employed a microarray approach using the Affymetrix Hugene 2.0 ST array chip followed by analysis with Ingenuity Pathways Analysis (IPA) software. In vitro studies showed that IL-15 decreased PCa cell motility at both concentrations. In vivo studies showed that IL-15 increased neutrophil infiltration, and the expression of adiponectin, desmin and alpha smooth muscle actin (α-sma) in the tumor tissue. Angiogenesis analysis, using CD31 immunohistochemistry, showed that IL-15 decreased the number of blood vessels. Gene expression analysis identified Cancer, Cell Death, Immune Response and Lipid Metabolism as the major diseases and functions altered in tumors treated with IL-15. This suggests that IL-15 causes inflammation and changes in stroma that can promote decreased tumor cell proliferation.

Integrin Inhibitors in Prostate Cancer

Prostate cancer (PCa) is the most frequently diagnosed cancer and the third highest cause of cancer-related deaths in men in the U.S. The development of chemotherapeutic agents that can bind PCa tumor cells with high specificity is critical in order to increase treatment effectiveness. Integrin receptors and their corresponding ligands have different expression patterns in PCa cells. They have been identified as promising targets to inhibit pathways involved in PCa progression. Currently, several compounds have proven to target specific integrins and their subunits in PCa cells. In this article, we review the role of integrins inhibitors in PCa and their potential as therapeutic targets for PCa treatments. We have discussed the following: natural compounds, monoclonal antibodies, statins, campothecins analog, aptamers, d-aminoacid, and snake venom. Recent studies have shown that their mechanisms of action result in decrease cell migration, cell invasion, cell proliferation, and metastasis of PCa cells.

Abstract 590: Andrographolide suppresses prostate cancer cell migration and alters the expression of vimentin, ZO-1 and MMP-11

Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL Prostate cancer is the most common cancer and the second cause of cancer-related deaths in males in the United States. The development of novel chemopreventive strategies may be extremely helpful to prevent and arrest the development of this neoplasm. Andrographolide, a labdane diterpenoid that is the main bioactive component of the medicinal plant Andrographis paniculata, has anti-inflammatory and anticarcinogenic properties. In this study, we sought to examine the effect of andrographolide on PC3 and LNCaP prostate cancer epithelial cells. Andrographolide significantly inhibited PC3 cell growth at a concentration of 10µM after 24 h of treatment (P < 0.001), and inhibited LNCaP cell growth at a concentration of 15 µM after 24 h of treatment (P < 0.001). Wound healing assay and boyden chamber experiments were performed in order to determine whether andrographolide had an effect on migration and invasion. When PC3 and LNCaP cells were treated with andrographolide the total number of migrating and invading cells was significantly reduced (P < 0.001) when compared to control. To study possible anti-migration and anti-invasion mechanisms of andrographolide on PC3 and LNCap cells, we examined the expression of various adhesion molecules. Andrograholide significantly decreased the expression of vimentin in PC3 and LNCap treated cells. However, PC3 and LNCaP cells treated with andrographolide had increased expression levels of occludin and ZO-1. In addition, immunofluorescence and western blot analysis showed that PC3 and LNCaP cells treated with andrographolide had decresased expression levels of MMP-11. Our results suggest that the anti-migration and anti-invasion effects of andrographolide in PC3 and LNCaP cells may be associated with alterations in the expression of occluding, ZO-1 and MMP-11. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 590. doi:1538-7445.AM2012-590

Cervicovaginal Fungi and Bacteria Associated With Cervical Intraepithelial Neoplasia and High-Risk Human Papillomavirus Infections in a Hispanic Population

The human cervicovaginal microbiota resides at an interface between the host and the environment and may affect susceptibility to disease. Puerto Rican women have high human papillomavirus (HPV) infection and cervical cancer rates. We hypothesized that the population structure of the cervicovaginal bacterial and fungal biota changed with cervical squamous intraepithelial lesions and HPV infections. DNA was extracted from cervix, introitus, and anal sites of 62 patients attending high-risk San Juan clinics. The 16S rRNA V4 region and ITS-2 fungal regions were amplified and sequenced using Illumina technology. HPV genotyping was determined by reverse hybridization with the HPV SPF10-LiPA25 kit. HPV prevalence was 84% of which ∼44% subjects were infected with high-risk HPV, ∼35% were co-infected with as many as 9 HPV types and ∼5% were infected with exclusively low-risk HPV types. HPV diversity did not change with cervical dysplasia. Cervical bacteria were more diverse in patients with CIN3 pre-cancerous lesions. We found enrichment of Atopobium vaginae and Gardnerella vaginalis in patients with CIN3 lesions. We found no significant bacterial biomarkers associated with HPV infections. Fungal diversity was significantly higher in cervical samples with high-risk HPV and introitus samples of patients with Atypical Squamous Cells of Undetermined Significance (ASCUS). Fungal biomarker signatures for vagina and cervix include Sporidiobolaceae and Sacharomyces for ASCUS, and Malassezia for high-risk HPV infections. Our combined data suggests that specific cervicovaginal bacterial and fungal populations are related to the host epithelial microenvironment, and could play roles in cervical dysplasia.

Abstract 5157: Preparation of curcumin analog nanoparticles and determination of their anticancer activity in prostate cancer

Prostate cancer is the most frequently diagnosed cancer and the second cause of cancer-related deaths in American men. Bioavailability of therapeutic agents is important for the treatment effectiveness. Previous studies have shown that the development of nanoparticles (NPs) drug delivery vehicles offers an opportunity for targeted drug delivery to tumor cells. Natural compounds such as curcumin have shown decreased bioavailability and stability when used as anticancer agents. Therefore, the purpose of this study is to synthesize the curcumin analog m-nitrochalcone (3bNchalc) encapsulated into poly (lactic-co-glycolic acid) (PLGA) NPs and determine the anticancer activity against prostate cancer cell lines. In our approach, 3bNChalc were encapsulated into PLGA NPs in the presence of PVA using the single emulsion-solvent evaporation method. 3bNChalc loading, encapsulation efficiency, and drug release was determined by spectrophotometric techniques. Physico-chemical properties such as zeta potential, particle size, polydispersity index (PDI), and morphology was measured using a combination of Dynamic Light Scattering (DLS) and Scanning Electron Microscopy (SEM). Cell viability and proliferation of PC3 and 22RV1 prostate cancer cell lines treated with 3bNchalc NPs and control were assessed using the MTS assay. 3bNchalc PLGA NPs were found to have a particle size of 250 nm and smooth spherical shape. In addition, zeta potential and mobility values revealed that 3bNChalc PLGA NPs are negatively charged and they tend to repel each other avoiding the tendency to flocculate. Moreover, 22RV1 cells treated with 3bNchalc PLGA NPs showed decreased in viability and proliferation when compared to control. We demonstrated that 3bNChalc was successfully encapsulated into PLGA NPs. In addition, our results showed that 3bNchalc PLGA NPs decreased the viability of 22RV1 prostate cancer cells when compared to control, suggesting that PLGA improves the delivery of 3bNChalc inside the cell. Results of our study will impact broadly the field by developing more effective and less toxic PLGA NPs based therapies. Citation Format: Maylein C. Juan-Rivera, Maria M. Sanchez-Vazquez, Gamalier Maldonado, Geovanny Ruiz, Noralejandra Vazquez, Christian Velez, Beatriz Zayas, Carlos Cabrera, Magaly Martinez-Ferrer, David Sanabria-Rios. Preparation of curcumin analog nanoparticles and determination of their anticancer activity in prostate 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 5157. doi:10.1158/1538-7445.AM2017-5157

Abstract 1537: The biological role of BAT-1 in prostate cancer

Prostate cancer is the most common cancer in men in the United States and is the second cause of cancer death in men. In Puerto Rico, prostate cancer is the most common type of cancer and the leading cause of cancer death in men. The current available biomarkers are unable to predict malignant outcomes such as recurrence. Thus, there is a critical demand for the development of innovative diagnostic and prognostic tools for the management of prostate cancer. In the current study, we evaluated the biological role of BAT-1 in prostate cancer. Preliminary data from patients who had prostate cancer recurrence identified that HLA-B associated transcript 1 (BAT-1) was down-regulated in patients with prostate cancer recurrence when compared with non-recurrent patients. We down-regulated BAT-1 in PC3 and 22RV1 prostate cancer cell lines using small interfering RNA (siRNA). In vitro assays were performed to measure de-differentiation, proliferation, apoptosis, migration and invasion. Flow cytometry analysis using the cell Viability/Annexin V cell staining kit showed that down-regulation of BAT-1 significantly increased the proliferation of PC3 cells (P Citation Format: Aileen M. Garcia-Vargas, Maylein C. Juan-Rivera, Milaris M. Sanchez-Cordero, Maria Sanchez-Vazquez, Krizia Rohena, Magaly Martinez-Ferrer. The biological role of BAT-1 in prostate cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1537.

Abstract 4640: Andrographolide modulates cell cycle, cell migration and tumor growth in prostate cancer

Prostate cancer is the most frequently diagnosed non-cutaneous cancer and is the second leading cause of cancer death in American men. Chemopreventive strategies have been tested to prevent the development of different types of cancer including prostate cancer. Andrographolide, a labdane diterpenoid that is the main bioactive component of the medicinal plant Andrographis paniculata, has been reported to have a wide range of biological activities including anticarcinogenic properties. In this study we aim to determine the role of Andrographolide in the progression of prostate cancer using in vitro and in vivo models. Andrographolide significantly inhibited PC3 cell growth at a concentration of 10μM after 24 h of treatment (P Citation Format: Ingrid Forestier-Roman, Maria Sanchez, Joseph Casillas, Krizia Rohena, Magaly Martinez-Ferrer. Andrographolide modulates cell cycle, cell migration and tumor growth in prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4640. doi:10.1158/1538-7445.AM2015-4640