Focal Segmental Glomerulosclerosis Complicating Therapy With Inotersen, an Antisense Oligonucleotide Inhibitor: A Case Report.

Inotersen is an antisense oligonucleotide inhibitor licensed for the treatment of polyneuropathy complicating hereditary transthyretin amyloidosis (ATTRv). Nephrotoxicity has been reported with inotersen, including progression to kidney failure. We describe what is to our knowledge the first reported case of inotersen-associated nephrotic syndrome secondary to focal segmental glomerulosclerosis (FSGS) and review the literature concerning inotersen-induced nephrotoxicity. We report a woman in her early 30s with ATTRv associated with the V50M transthyretin (TTR) variant, who presented with nephrotic syndrome 7 months after commencement of inotersen. Renal histology demonstrated FSGS and scanty glomerular amyloid deposition. Discontinuation of inotersen alone resulted in complete clinical and biochemical resolution of nephrotic syndrome. Inotersen is associated with significant nephrotoxicity. In the phase 3 NEURO-TTR clinical trial, 3% of patients in the treatment arm developed a crescentic glomerulonephritis. All affected patients carried the V50M TTR variant, which is known to be associated with renal amyloid deposition. This case adds to the spectrum of kidney disease associated with inotersen and indicates that discontinuation of the drug alone may result in resolution of renal complications without additional immunosuppression. Monitoring of kidney function is essential in patients with ATTRv receiving inotersen, particularly if there is evidence of existing renal amyloid.

The effect of admission and pre-admission serum creatinine as baseline to assess incidence and outcomes of acute kidney injury in acute medical admissions.

BACKGROUND: Acute kidney injury (AKI) in hospital-admitted patients is a common complication associated with increased mortality. The diagnosis of AKI relies on the ascertainment of peak increase in serum creatinine (SCr). This study evaluated the incidence of AKI using the increase from mean 7-365 days pre-admission (AKIpre) and admission (AKIadm) SCr levels, and examined the associations of AKI and changes in SCr levels with all-cause mortality. METHODS: A total of 2436 patients admitted to a tertiary hospital were recruited and followed-up for a median of 47.70 (interquartile range 18.20) months. AKI incidence and severity were defined according to the Kidney Disease: Improving Global Outcomes-AKI Guidelines. Follow-up data were collected from the Hospital Episode Statistics and Office of National Statistics. Mortality was evaluated during a short- (30 days), mid- (1 year) and long-term (4 years) period. RESULTS: No difference in the AKI rates using AKIpre and AKIadm (12.5% versus 12.2%; P = 0.695) or in the AKI severity (P = 0.261) was evident. Agreement between the two definitions was modest (Kappa-statistic = 0.596, P < 0.001). Patients with AKIpre or AKIadm had increased all-cause mortality compared with those without AKI during all follow-up periods. In fully adjusted regression analysis, AKIpre [hazard ratio (HR) = 2.226, 95% confidence interval (CI) 1.140-4.347; P = 0.027] and AKIadm (HR = 2.105, 95% CI 1.090-4.064; P = 0.027) remained associated with 30-day mortality. Results for the 1- and 4-year periods were similar. Increases of >4.00 µmol/L and >6.06% from pre-admission or >6.00 µmol/L and >17.24% from admission SCr levels presented increased mortality risk during follow-up. CONCLUSIONS: Use of admission or pre-admission SCr provides similar incidence rates, but they diagnose different sets of patients. Even minor increases in SCr, below those required for the classification of AKI, were associated with increased mortality. These findings can help the clinicians to identify patients at higher risk for adverse outcomes.

Incidence and impact on outcomes of acute kidney injury after a stroke: a systematic review and meta-analysis.

BACKGROUND: Patients with chronic kidney disease have worse outcomes after stroke. However, the burden of acute kidney injury after stroke has not been extensively investigated. METHODS: We used MEDLINE and Embase to conduct a systematic review and meta-analysis of published studies that provided data on the risk of AKI and outcomes in adults after ischemic and hemorrhagic stroke. Pooled incidence was examined using the Stuart-Ord method in a DerSimonian-Laird model. Pooled Odds Ratios and 95% confidence intervals were calculated for outcomes using a random effects model. This review was registered with PROSPERO (CRD42017064588). RESULTS: Eight studies were included, five from the United States, representing 99.9% of included patients. Three studies used established acute kidney injury criteria based on creatinine values to define acute kidney injury and five used International Classification of Diseases coding definitions. Overall pooled incidence was 9.61% (95% confidence interval 8.33-10.98). Incidence for studies using creatinine definitions was 19.51% (95% confidence interval 12.75-27.32%) and for studies using coding definitions 4.63% (95% confidence interval 3.65-5.72%). Heterogeneity was high throughout. Mortality in stroke patients who sustained acute kidney injury was increased (Odds Ratio 2.45; 95% confidence interval 1.47-4.10). Three studies reported risk factors for acute kidney injury. There was sparse information on other outcomes. CONCLUSIONS: Mortality in stroke patients who develop acute kidney injury is significantly increased. However the reported incidence of AKI after stroke varies widely and is underestimated using coding definitions. Larger international studies are required to identify potentially preventable factors to reduce acute kidney injury after stroke and improve outcomes.

Fracture risk and mortality post-kidney transplantation.

BACKGROUND: Fractures are associated with high morbidity and economic costs. There is a paucity of information on fractures after kidney transplantation outside the United States. METHODS: Data were obtained from the Hospital Episode Statistics database on kidney transplants performed in England between 2001 and 2013 and post-transplant fracture-related hospitalization. Mortality data were obtained from the Office for National Statistics. RESULTS: In total, 21 769 first kidney transplant procedures were analyzed with 112 512 patient-years follow-up. Overall, 836 (3.8%) kidney allograft recipients developed a fracture requiring hospitalization. Event rate was 9.99 for any fracture and 1.54 for a hip fracture per 1000 patient-years. Accounting for the competing risk of mortality, increasing age, female gender, white ethnicity, and a history of pre-transplant diabetes mellitus or previous fracture were associated with increased fracture risk post-kidney transplantation. Death occurred in 2407 (11.1%) kidney allograft recipients, with 173 deaths occurring post-fracture. In an extended Cox model, hip fracture as a time-varying factor was independently associated with an increased risk of death (hazard ratio, 3.288; 95% confidence intervals, 2.513-4.301; p < 0.001). CONCLUSIONS: Fracture rates in English kidney transplant recipients are lower than previously reported in US cohorts. Sustaining a hip fracture is associated with an increased mortality risk. Our results can be used to power future fracture prevention trials.

On the complex relationship between resilience and hair cortisol levels in adolescence despite parental physical abuse: a fourth wave of resilience research.

Introduction: To understand the family's role in adolescents' mental health development and the connection to neurodevelopmental disorders related to experienced parental physical abuse, we first explored resilience pathways longitudinally and secondly, connected the identified patterns to adolescents' hair cortisol levels that are rooted in the hypothalamic-pituitary-adrenal axis as the main stress response system and connected brain structure alterations. Methods: We analyzed longitudinal online questionnaire data for three consecutive high school years (from seventh to ninth grade) and four survey waves from a representative sample of n = 1609 high school students in Switzerland on violence-resilience pathways. Furthermore, we collected students' hair samples from a subsample of n = 229 at survey wave 4. About 30% of the participating adolescents had been physically abused by their parents. Out of the overall sample, we drew a subsample of adolescents with parental abuse experiences (survey wave 1 n = 509; survey wave 2 n = 506; survey wave 3 n = 561; survey wave 4 n = 560). Results: Despite the odds, about 20-30% of adolescents who have experienced parental physical abuse escaped the family violence cycle and can be called resilient. By applying a person-oriented analytical approach via latent class and transition analysis, we longitudinally identified and compared four distinct violence-resilience patterns. We identified violence resilience as a multidimensional latent construct, which includes hedonic and eudaimonic protective and risk indicators. Because resilience should not solely be operationalized based on the lack of psychopathology, our latent construct included both feeling good (hedonic indicators such as high levels of self-esteem and low levels of depression/anxiety and dissociation) and doing well (eudaimonic indicators such as high levels of self-determination and self-efficacy as well as low levels of aggression toward peers). Discussion: The present study confirmed that higher cortisol levels significantly relate to the comorbid pattern (internalizing and externalizing symptoms), and further confirmed the presence of lasting alterations in brain structures. In this way, we corroborated the insight that when studying the resilience pathways and trajectories of abused adolescents, biological markers such as hair cortisol significantly enhance and deepen the understanding of the longitudinal mechanisms of psychological markers (e.g., self-determination, self-esteem, self-efficacy) that are commonly applied in questionnaires.

Evidence of neurotrophic events due to peritoneal endometriotic lesions.

To investigate the neurotrophic properties of endometriosis, as well as the involvement of neurotrophic factors in the development of chronic pelvic pain in patients with endometriosis, we performed a prospective clinical study. The presence of neurotrophins was investigated in the peritoneal fluid (PF) of patients with peritoneal endometriotic lesions or adenomyosis, as well as from women with non-endometriotic adhesions and from women without endometriosis/adenomyosis/adhesions. The PF from patients with peritoneal endometriotic lesions was divided in three groups: asymptomatic endometriosis, minimal pain and severe pain. PF from patients with adenomyosis or with non-endometriotic adhesions and the control group were divided in patients without pain and with pain. Neurotrophin expression in PF was analyzed using Elisa and the neuronal growth assay with cultured chicken sensory ganglia (dorsal-root-ganglia, DRG) and sympathetic ganglia. PF from women with peritoneal endometriotic lesions overexpress nerve growth factor (NGF) and neurotrophin-3 (NT-3), but not brain derived neurotrophic factor (BDNF), whereas the PF of women with adenomyosis or adhesions seems to express normal amounts of these factors. Neurotrophin expression did not differ among the pain groups. Furthermore, the PF from patients with peritoneal endometriotic lesions induced a strong sensory and a marginal sympathetic neurite outgrowth, while the PF from women with adenomyosis and non-endometriotic adhesions induced an outgrowth similar to the control group. The induced neurite outgrowth could only be inhibited in DRG incubated with peritoneal endometriotic lesions. Interestingly, the outgrowth of sympathetic ganglia was inhibited in all studied groups. The present study suggests that only peritoneal endometriotic lesions lead to an increased release of NGF and NT-3 into the PF and that NGF modulates the nerve fiber growth in endometriosis.

Neuroimmunomodulatory alterations in non-lesional peritoneum close to peritoneal endometriosis.

OBJECTIVES: An imbalance in the ratio of sensory to sympathetic nerve fibre (NF) density in peritoneal endometriotic lesions (pEL) has recently been demonstrated and leads to the assumption that this preponderance of the sensory pro-inflammatory milieu is a major cause of pain in endometriosis. Therefore, the density of sensory and sympathetic NFs was determined in distal unaffected peritoneum of endometriosis patients to be able to detect possible alterations in unaffected peritoneum. METHODS: In serial pEL sections (n = 40), lesional and matching unaffected peritoneum as well as healthy peritoneum (HP) from patients without endometriosis (n = 15) were immunohistochemically analysed to identify protein gene product 9.5-, substance P- and tyrosine hydroxylase-positive NFs (intact, sensory and sympathetic NFs, respectively). In addition, the amount of immune cell infiltrates and the expression of nerve growth factor (NGF) and interleukin (IL)-1ß in nerves of peritoneal endometriotic specimens were compared to those in the HP. RESULTS: The overall NF density in the non-lesional, unaffected peritoneum of endometriosis patients is significantly reduced in comparison to both HP and pEL, while sensory NFs remain the same; the sympathetic NF density is significantly decreased compared to HP, but is still higher than the density close to the pEL. Immune cell infiltrates as well as NGF and IL-1ß expression in nerves is significantly elevated in distal unaffected peritoneum in comparison to HP. CONCLUSION: The altered NF density in the non-lesional, unaffected peritoneum of endometriosis patients suggests new aspects in the understanding of the development of endometriosis and pain management in endometriosis.

Integrating ayurvedic medicine into cancer research programs part 2: Ayurvedic herbs and research opportunities.

The aim of this two-part review in this issue is to provide some basic perspectives from Ayurveda, the traditional medicine of India, and to discuss how current research methodologies may be used to shed light on mechanisms of Ayurvedic treatments to support cancer care and prevention. It addresses some of the challenges for scientific validation of Ayurvedic herbal compounds, protocols, and modalities in four areas. Part 1 [1] has reviewed Ayurvedic theories and applications of body constitution (Prakriti), digestion (Agni and Ama) and mind-body-spirit health in relation to cancer. Here in Part 2, the focus is on preclinical and clinical research of Ayurvedic botanical herbs, with a review of pertinent literature on three selected herbs, Curcumin, Ashwagandha, and Triphala. A discussion of the challenges and possibilities of research in Ayurveda is offered to guide the development of translational research programs. Ayurvedic modalities are not intended as a substitute for allopathic treatments of cancer but as an integrative component for prevention and restoration of strength and immunity.

Integrating ayurvedic medicine into cancer research programs part 1: Ayurveda background and applications.

Integration of Ayurveda into our current health care research programs is critical to making progress in global wellness and in disease prevention and control, especially for cancer. Ayurveda promotes restoration of the innate healing mechanisms existing in the body for optimal immunity, resilience, and health. Ayurveda also has an abundant resource of botanical products containing diverse pharmaco-active ingredients and millennia of experience of clinical applications for health benefits. But there is a lack of evidence-based research to demonstrate its efficacy and potential. This 2-part review is written from the perspective of a western-trained biomedical scientist and student of Ayurveda. It aims to educate research scientist peers about the opportunities and challenges for scientific validation of Ayurvedic herbal compounds, protocols, and modalities and inspire more research in this area. Part 1 will review several aspects of Ayurveda including principles of body constitution (Prakriti), digestion (Agni and Ama) and mind-body health, in relation to cancer. Part 2 [1] will focus on Ayurvedic botanical resources used for cancer and research studies will be discussed on selected herbal compounds. Research gaps and opportunities will be identified to guide development of research programs to validate safety and efficacy of these therapies. Importantly, the use of Ayurvedic modalities is not intended to substitute for allopathic treatments for cancer but as an integrative component for prevention and restoration of strength and immunity post treatment.

Imbalance between sympathetic and sensory innervation in peritoneal endometriosis.

To investigate possible mechanisms of pain pathophysiology in patients with peritoneal endometriosis, a clinical study on sensory and sympathetic nerve fibre sprouting in endometriosis was performed. Peritoneal lesions (n=40) and healthy peritoneum (n=12) were immunostained and analysed with anti-protein gene product 9.5 (PGP 9.5), anti-substance P (SP) and anti-tyrosine hydroxylase (TH), specific markers for intact nerve fibres, sensory nerve fibres and sympathetic nerve fibres, respectively, to identify the ratio of sympathetic and sensory nerve fibres. In addition, immune cell infiltrates in peritoneal endometriotic lesions were analysed and the nerve growth factor (NGF) and interleukin (IL)-1ß expression was correlate with the nerve fibre density. Peritoneal fluids from patients with endometriosis (n=40) and without endometriosis (n=20) were used for the in vitro neuronal growth assay. Cultured chicken dorsal root ganglia (DRG) and sympathetic ganglia were stained with anti-growth associated protein 43 (anti-GAP 43), anti-SP and anti-TH. We could detect an increased sensory and decreased sympathetic nerve fibres density in peritoneal lesions compared to healthy peritoneum. Peritoneal fluids of patients with endometriosis compared to patients without endometriosis induced an increased sprouting of sensory neurites from DRG and decreased neurite outgrowth from sympathetic ganglia. In conclusion, this study demonstrates an imbalance between sympathetic and sensory nerve fibres in peritoneal endometriosis, as well as an altered modulation of peritoneal fluids from patients with endometriosis on sympathetic and sensory innervation which might directly be involved in the maintenance of inflammation and pain.

Transforming growth factor ß1 increase of hydroxysteroid dehydrogenase proteins is partly suppressed by red clover isoflavones in human primary prostate cancer-derived stromal cells.

Transforming growth factor ß1 (TGF-ß1) increases dehydro-epiandrosterone (DHEA) metabolism to androgens and prostate-specific antigen (PSA) in a prostate tissue model where stromal (6S) cells and epithelial (LAPC-4) cells are cocultured. Red clover (RC) isoflavones inhibits transforming growth factor (TGF)-ß-induced androgenicity. Mechanisms controlling those activities were explored. Three hydroxysteroid dehydrogenases (HSDs), 3ß-HSD, HSD-17ß1 and HSD-17ß5 involved in metabolizing DHEA to testosterone (TESTO) were investigated. Individual depletion of HSDs in 6S cells significantly reduced TGF-ß1/DHEA-induced PSA in LAPC-4 cells in cocultures. Monomer amounts of 3ß-HSD were similar without or with TGF-ß1 in both cell types but aggregates of 3ß-HSD in 6S cells were much higher than those in LAPC-4 cells and were upregulated by TGFß in 6S cells. Basal and TGF-ß1-treated levels of HSD-17ß1 and HSD-17ß5 in LAPC-4 cells were significantly lower than in 6S cells, whereas levels of HSD-17ß1 but not HSD-17ß5 were TGFß inducible. 6S cell HSD genes expression induced by TGFß or androgen signaling was insignificant to contribute TGF-ß1/DHEA-upregulated protein levels of HSDs. RC decreased TGF-ß1- upregulation of aggregates of 3ß-HSD but not HSD-17ß1. Depletion of TGFß receptors (TGFß Rs) reduced TGF-ß1/DHEA-upregulated HSDs and TESTO. Immunoprecipitation studies demonstrated that TGF-ß1 disrupted associations of TGFß Rs/HSDs aggregates, whereas RC suppressed the dissociations of aggregates of 3ß-HSD but not HSD-17ß1 from the receptors. Given that TGFß Rs are recycled with or without ligand, TGF-ß1-induced disassociation of the HSDs from TGFß Rs may increase stability and activity of the HSDs. These data suggest a pathway connecting overproduction of TGFß with increased PSA in prostate cancer.

Androgen-induced PSA expression requires not only activation of AR but also endogenous IGF-I or IGF-I/PI3K/Akt signaling in human prostate cancer epithelial cells.

BACKGROUND: Prostate cancer (PrCa) risk is positively associated with levels of insulin-like growth factor I (IGF-I) and prostate specific antigen (PSA), both androgen receptor (AR) signaling target genes in PrCa cells. Although activated AR is required for androgen-induction of expression of both genes, effects of the IGF-I signaling pathways on the androgen-induction of PSA have not been studied. METHODS: Human prostate stromal and epithelial cancer cells were treated alone or in coculture with steroid hormone and/or inhibitors. Gene or protein expression was analyzed by real time RT-PCR or Western blotting of lysates, nuclear extracts, or immunoprecipitated products. RESULTS: In PrCa epithelial cells, endogenous IGF-I, significantly induced by R1881, was required for R1881-induction of PSA. Increased IGF-I correlated with accumulation of cytoplasmic dephospho ß-catenin (CPDP ß-catenin), a co-activator of AR signaling. Exogenous IGF-I enhanced R1881-induced PSA and accumulation of CPDP ß-catenin in LAPC-4 cells. Functional depletion of IGF-I or IGF-I receptor diminished PSA induction. Induction of IGF-I reached a plateau while PSA consecutively increased. Inhibiting PI3K abolished R1881-induced Akt phosphorylation, CPDP and nuclear ß-catenin and nuclear association of AR/ß-catenin, consequently abrogating R1881-induced expression of IGF-I and/or PSA. CONCLUSIONS: By integrating androgen, IGF-I and ß-catenin signaling pathways, these data reveal that androgen-induced PSA expression requires activation of AR and endogenous IGF-I or IGF-I/PI3K/Akt signaling, suggesting a positive feedback cycle for increased production of PSA associated with PrCa.

Motor, sensitive, and vegetative recovery in rats with compressive spinal-cord injury after combined treatment with erythropoietin and whole-body vibration.

BACKGROUND: Physical therapy with whole body vibration (WBV) following compressive spinal cord injury (SCI) in rats restores density of perisomatic synapses, improves body weight support and leads to a better bladder function. The purpose of the study was to determine whether the combined treatment with WBV plus erythropoietin (EPO) would further improve motor, sensory and vegetative functions after SCI in rats. METHODS: Severe compressive SCI at low thoracic level was followed by a single i.p. injection of 2,5µg (250 IU) human recombinant EPO. Physical therapy with WBV started on 14th day after injury and continued over a 12-week post injury period. Locomotor recovery, sensitivity tests and urinary bladder scores were analysed at 1, 3, 6, 9, and 12 weeks after SCI. The closing morphological measurements included lesion volume and numbers of axons in the preserved perilesional neural tissue bridges (PNTB). RESULTS: Assessment of motor performance sensitivity and bladder function revealed no significant effects of EPO when compared to the control treatments. EPO treatment neither reduced the lesion volume, nor increased the number of axons in PNTB. CONCLUSIONS: The combination of WBV + EPO exerts no positive effects on hind limbs motor performance and bladder function after compressive SCI in rats.

Impact of elevated N input on soil N cycling and losses in old-growth lowland and montane forests in Panama.

Nitrogen deposition is projected to increase rapidly in tropical ecosystems, but changes in soil-N-cycling processes in tropical ecosystems under elevated N input are less well understood. We used N-addition experiments to achieve N-enriched conditions in mixed-species, lowland and montane forests in Panama. Our objectives were to (1) assess changes in soil mineral N production (gross rates of N mineralization and nitrification) and retention (microbial immobilization and rapid reactions to organic N) during 1- and 9-yr N additions in the lowland forest and during 1-yr N addition in the montane forest and (2) relate these changes to N leaching and N-oxide emissions. In the old-growth lowland forest located on an Inceptisol, with high base saturation and net primary production not limited by N, there was no immediate effect of first-year N addition on gross rates of mineral-N production and N-oxide emissions. Changes in soil-N processes were only apparent in chronic (9 yr) N-addition plots: gross N mineralization and nitrification rates, NO3- leaching, and N-oxide emissions increased, while microbial biomass and NH4+ immobilization rates decreased compared to the control. Increased mineral-N production under chronic N addition was paralleled by increased substrate quality (e.g., reduced C:N ratios of litterfall), while the decrease in microbial biomass was possibly due to an increase in soil acidity. An increase in N losses was reflected in the increase in 15N signatures of litterfall under chronic N addition. In contrast, the old-growth montane forest located on an Andisol, with low base saturation and aboveground net primary production limited by N, reacted to first-year N addition with increases in gross rates of mineral-N production, microbial biomass, NO3- leaching, and N-oxide emissions compared to the control. The increased N-oxide emissions were attributed to increased nitrification activity in the organic layer, and the high NO3- availability combined with the high rainfall on this sandy loam soil facilitated the instantaneous increase in NO3-leaching. These results suggest that soil type, presence of an organic layer, changes in soil-N cycling, and hydrological properties are more important indicators than vegetation as an N sink on how tropical forests respond to elevated N input.

Acute kidney injury calculated using admission serum creatinine underestimates 30-day and 1-year mortality after acute stroke.

BACKGROUND: Acute kidney injury (AKI) diagnosis requires ascertainment of change from a known baseline. Although pre-admission serum creatinine (SCr) is recommended, to date, all studies of AKI in acute stroke have used the first SCr on admission. METHODS: All patients admitted with an acute stroke to an emergency hospital were recruited. We compared use of pre-admission SCr with admission SCr to diagnose AKI. Regression analyses were used to identify risk factors for 30-day and 1-year mortality, respectively. RESULTS: A total of 1354 patients were recruited from December 2012 to September 2015. Incidence of AKI was 18.7 and 19.9% using pre-admission SCr and admission SCr, respectively. Diagnosis of AKI was associated with significantly increased 30-day and 1-year mortality. Diagnosis of AKI using pre-admission SCr had a stronger relationship with both 30-day and 1-year mortality. In 443 patients with a pre-admission SCr and at least two SCr during admission, AKI diagnosed using pre-admission SCr had a stronger relationship than AKI diagnosed using admission SCr with 30-day mortality [odds ratio (OR) = 2.64; 95% confidence interval (CI) 1.36-5.12; P = 0.004 versus OR = 2.10; 95% CI 1.09-4.03; P = 0.026] and 1-year mortality [hazard ratio (HR) = 1.90, 95% CI 1.32-2.76; P = 0.001 versus HR = 1.47; 95% CI 1.01-2.15; P = 0.046] in fully adjusted models. CONCLUSIONS: AKI after stroke is common and is associated with increased 30-day and 1-year mortality. Using first SCr on admission gives a comparable AKI incidence to pre-admission SCr, but underestimates 30-day and 1-year mortality risk.

Tubulin Tyrosine Ligase Like 4 (TTLL4) overexpression in breast cancer cells is associated with brain metastasis and alters exosome biogenesis.

BACKGROUND: The survival rate is poor in breast cancer patients with brain metastases. Thus, new concepts for therapeutic approaches are required. During metastasis, the cytoskeleton of cancer cells is highly dynamic and therefore cytoskeleton-associated proteins are interesting targets for tumour therapy. METHODS: Screening for genes showing a significant correlation with brain metastasis formation was performed based on microarray data from breast cancer patients with long-term follow up information. Validation of the most interesting target was performed by MTT-, Scratch- and Transwell-assay. In addition, intracellular trafficking was analyzed by live-cell imaging for secretory vesicles, early endosomes and multiple vesicular bodies (MVB) generating extracellular vesicles (EVs). EVs were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), Western blotting, mass spectrometry, and ingenuity pathway analysis (IPA). Effect of EVs on the blood-brain-barrier (BBB) was examined by incubating endothelial cells of the BBB (hCMEC/D3) with EVs, and permeability as well as adhesion of breast cancer cells were analyzed. Clinical data of a breast cancer cohort was evaluated by χ2-tests, Kaplan-Meier-Analysis, and log-rank tests while for experimental data Student's T-test was performed. RESULTS: Among those genes exhibiting a significant association with cerebral metastasis development, the only gene coding for a cytoskeleton-associated protein was Tubulin Tyrosine Ligase Like 4 (TTLL4). Overexpression of TTLL4 (TTLL4plus) in MDA-MB231 and MDA-MB468 breast cancer cells (TTLL4plus cells) significantly increased polyglutamylation of ß-tubulin. Moreover, trafficking of secretory vesicles and MVBs was increased in TTLL4plus cells. EVs derived from TTLL4plus cells promote adhesion of MDA-MB231 and MDA-MB468 cells to hCMEC/D3 cells and increase permeability of hCMEC/D3 cell layer. CONCLUSIONS: These data suggest that TTLL4-mediated microtubule polyglutamylation alters exosome homeostasis by regulating trafficking of MVBs. The TTLL4plus-derived EVs may provide a pre-metastatic niche for breast cancer cells by manipulating endothelial cells of the BBB.

DHEA metabolism in prostate: For better or worse?

Dehydroepiandrosterone (DHEA) is commonly used in the USA as a nutritional supplement for antiaging, metabolic support or other uses. Investigations into understanding the effects of DHEA on human prostate cancer progression have posed more questions than answers and highlight the importance of communications between stromal and epithelial tuoitiuot elements within the prostate that contribute to the regulation of DHEA metabolism. Intracrine metabolism of DHEA to androgens (A) and/or estrogens (E) may occur in one cell compartment (stromal) which may release paracrine hormones or growth/inhibitory factors to the epithelial cells. Alternatively no metabolism of DHEA may occur, resulting in no harmful consequences of high levels of DHEA in prostate tissues. We herein review the tissue components involved and interactions with the prohormone, DHEA and/or resulting metabolites, including dihydrotestosterone (DHT) or 17beta-estradiol (E(2)) in an in vitro model of endocrine-immune-paracrine interactions within the prostate. This work raises questions and hypotheses concerning the role of DHEA in prostate in normal tissues, vs. preneoplastic tissues.

The Role of Lineage Plasticity in Prostate Cancer Therapy Resistance.

Lineage plasticity has emerged as an important mechanism of treatment resistance in prostate cancer. Treatment-refractory prostate cancers are increasingly associated with loss of luminal prostate markers, and in many cases induction of developmental programs, stem cell-like phenotypes, and neuroendocrine/neuronal features. Clinically, lineage plasticity may manifest as low PSA progression, resistance to androgen receptor (AR) pathway inhibitors, and sometimes small cell/neuroendocrine pathologic features observed on metastatic biopsy. This mechanism is not restricted to prostate cancer as other malignancies also demonstrate lineage plasticity during resistance to targeted therapies. At present, there is no established therapeutic approach for patients with advanced prostate cancer developing lineage plasticity or small cell neuroendocrine prostate cancer (NEPC) due to knowledge gaps in the underlying biology. Few clinical trials address questions in this space, and the outlook for patients remains poor. To move forward, urgently needed are: (i) a fundamental understanding of how lineage plasticity occurs and how it can best be defined; (ii) the temporal contribution and cooperation of emerging drivers; (iii) preclinical models that recapitulate biology of the disease and the recognized phenotypes; (iv) identification of therapeutic targets; and (v) novel trial designs dedicated to the entity as it is defined. This Perspective represents a consensus arising from the NCI Workshop on Lineage Plasticity and Androgen Receptor-Independent Prostate Cancer. We focus on the critical questions underlying lineage plasticity and AR-independent prostate cancer, outline knowledge and resource gaps, and identify strategies to facilitate future collaborative clinical translational and basic studies in this space.

Lycopene inhibits IGF-I signal transduction and growth in normal prostate epithelial cells by decreasing DHT-modulated IGF-I production in co-cultured reactive stromal cells.

Prostate stromal and epithelial cell communication is important in prostate functioning and cancer development. Primary human stromal cells from normal prostate stromal cells (PRSC) maintain a smooth muscle phenotype, whereas those from prostate cancer (6S) display reactive and fibroblastic characteristics. Dihydrotestosterone (DHT) stimulates insulin-like growth factor-I (IGF-I) production by 6S but not PSRC cells. Effects of reactive versus normal stroma on normal human prostate epithelial (NPE or PREC) cells are poorly understood. We co-cultured NPE plus 6S or PRSC cells to compare influences of different stromal cells on normal epithelium. Because NPE and PREC cells lose androgen receptor (AR) expression in culture, DHT effects must be modulated by associated stromal cells. When treated with camptothecin (CM), NPE cells, alone and in stromal co-cultures, displayed a dose-dependent increase in DNA fragmentation. NPE/6S co-cultures exhibited reduced CM-induced cell death with exposure to DHT, whereas NPE/PRSC co-cultures exhibited CM-induced cell death regardless of DHT treatment. DHT blocked CM-induced, IGF-I-mediated, NPE death in co-cultured NPE/6S cells without, but not with, added anti-IGF-I and anti-IGF-R antibodies. Lycopene consumption is inversely related to human prostate cancer risk and inhibits IGF-I and androgen signaling in rat prostate cancer. In this study, lycopene, in dietary concentrations, reversed DHT effects of 6S cells on NPE cell death, decreased 6S cell IGF-I production by reducing AR and beta-catenin nuclear localization and inhibited IGF-I-stimulated NPE and PREC growth, perhaps by attenuating IGF-I's effects on serine phosphorylation of Akt and GSK3beta and tyrosine phosphorylation of GSK3. This study expands the understanding of the preventive mechanisms of lycopene in prostate cancer.