Community Voice in Cross-Sector Alignment: Concepts and Strategies from a Scoping Review of the Health Collaboration Literature

BackgroundHealth care access is an important driver of population health, and factors beyond health care also drive health outcomes. Recognizing the importance of the social determinants of health (SDOH), different actors in the health care, public health, and social service sectors are increasingly collaborating to improve health outcomes in communities. To support such collaboration, the Robert Wood Johnson Foundation developed a cross-sector alignment theory of change. According to the cross-sector alignment theory of change, community voice is critical for helping collaboratives address community health needs. Yet research on health collaboratives offers mixed guidance on how community voice should be understood and which community voice strategies are most effective.MethodsThis study addresses a gap in the literature with a systematic scoping review of research on health-oriented cross-sector collaboration and community voice. By scanning key academic journals, searching three academic databases, and obtaining documents from across our professional networks, we identified 36 documents that address community voice in health collaboratives.ResultsThe review reveals several conceptions of community voice and a range of community voice strategies. We find that community voice strategies fall on a spectrum between two broad types of approaches: active and passive. These vary not only in the level of power shared between communities and collaborators, but also in the level of involvement required from the community, and this in turn has important implications for community collaboration strategies. We also find that while most strategies are discussed in the context of short-term collaboration, many also lend themselves to adoption in the context of sustainable collaboration and, ultimately, cross-sector alignment.ConclusionThis review provides a characterization and conceptualization of community voice in health-oriented collaborations that provides a new theoretical basis for future research. Passive and active community voice strategies can be studied in more detail for their expected impact on health outcomes and disparities. Increased attention to active community voice and the resources it requires can help practitioners achieve improved health outcomes and researchers understand the pathways to health improvement through collaboration

Polyphenols from the Mediterranean herb rosemary (Rosmarinus officinalis) for prostate cancer

The Mediterranean diet is rich in fruits and vegetables and has been associated with a variety of health benefits including cancer prevention. One aspect of the diet that has not received enough attention is Mediterranean herbs. Specifically, rosemary and its polyphenolic diterpenes (carnosic acid and carnosol) are known to possess anti-oxidant activity that may be beneficial for cancer control. Herein, we describe the in vitro and in vivo studies carried out towards understanding the molecular mechanisms of carnosic acid and carnosol leading to inhibition of prostate cancer. The reported findings suggest that these polyphenols target multiple signaling pathways involved in cell cycle modulation and apoptosis. Further work is required to understand its potential for health promotion and potential drug discovery for prostate cancer chemoprevention

α-Mangostin Promotes In Vitro and In Vivo Degradation of Androgen Receptor and AR-V7 Splice Variant in Prostate Cancer Cells

A major limitation of current prostate cancer pharmacotherapy approaches is the inability of these compounds to target androgen receptor variants or mutants that develop during prostate cancer progression. The demand for novel therapeutics to prevent, slow, and treat prostate cancer is significant because FDA approved anti-androgens are associated with adverse events and can eventually drive drug-resistant prostate cancer. This study evaluated α-mangostin for its novel ability to degrade the androgen receptor and androgen receptor variants. α-Mangostin is one of more than 70 isoprenylated xanthones isolated from Garcinia mangostana that we have been evaluating for their anticancer potential. Prostate cancer cells treated with α-mangostin exhibited decreased levels of wild-type and mutated androgen receptors. Immunoblot, immunoprecipitation, and transfection experiments demonstrated that the androgen receptor was ubiquitinated and subsequently degraded via the proteasome, which we hypothesize occurs with the assistance of BiP, an ER chaperone protein that we have shown to associate with the androgen receptor. We also evaluated α-mangostin for its antitumor activity and promotion of androgen receptor degradation in vivo. In summary, our study demonstrates that androgen receptor degradation occurs through the novel activation of BiP and suggests a new therapeutic approach for prostate cancer

Development of drug-inducible CRISPR-Cas9 systems for large-scale functional screening

Abstract Background Large-scale genetic screening using CRISPR-Cas9 technology has emerged as a powerful approach to uncover and validate gene functions. The ability to control the timing of genetic perturbation during CRISPR screens will facilitate precise dissection of dynamic and complex biological processes. Here, we report the optimization of a drug-inducible CRISPR-Cas9 system that allows high-throughput gene interrogation with a temporal control. Results We designed multiple drug-inducible sgRNA expression vectors and measured their activities using an EGFP gene disruption assay in 11 human and mouse cell lines. The optimal design allows for a tight and inducible control of gene knockout in vitro, and in vivo during a seven-week-long experiment following hematopoietic reconstitution in mice. We next performed parallel genome-wide loss-of-function screens using the inducible and constitutive CRISPR-Cas9 systems. In proliferation-based dropout screens, these two approaches have similar performance in discriminating essential and nonessential genes. In a more challenging phenotypic assay that requires cytokine stimulation and cell staining, we observed similar sensitivity of the constitutive and drug-induced screening approaches in detecting known hits. Importantly, we demonstrate minimal leakiness of our inducible CRISPR screening platforms in the absence of chemical inducers in large-scale settings. Conclusions In this study, we have developed a drug-inducible CRISPR-Cas9 system that shows high cleavage efficiency upon induction but low background activity. Using this system, we have achieved inducible gene disruption in a wide range of cell types both in vitro and in vivo. For the first time, we present a systematic side-by-side comparison of constitutive and drug-inducible CRISPR-Cas9 platforms in large-scale functional screens. We demonstrate the tightness and efficiency of our drug-inducible CRISPR-Cas9 system in genome-wide pooled screening. Our design increases the versatility of CRISPR-based genetic screening and represents a significant upgrade on existing functional genomics toolbox

Rosemary (Rosmarinus officinalis) extract modulates CHOP/GADD153 to promote androgen receptor degradation and decreases xenograft tumor growth.

The Mediterranean diet has long been attributed to preventing or delaying the onset of cardiovascular disease, diabetes and various solid organ cancers. In this particular study, a rosemary extract standardized to carnosic acid was evaluated for its potential in disrupting the endoplasmic reticulum machinery to decrease the viability of prostate cancer cells and promote degradation of the androgen receptor. Two human prostate cancer cell lines, 22Rv1 and LNCaP, and prostate epithelial cells procured from two different patients undergoing radical prostatectomy were treated with standardized rosemary extract and evaluated by flow cytometry, MTT, BrdU, Western blot and fluorescent microscopy. A significant modulation of endoplasmic reticulum stress proteins was observed in cancer cells while normal prostate epithelial cells did not undergo endoplasmic reticulum stress. This biphasic response suggests that standardized rosemary extract may preferentially target cancer cells as opposed to "normal" cells. Furthermore, we observed standardized rosemary extract to decrease androgen receptor expression that appears to be regulated by the expression of CHOP/GADD153. Using a xenograft tumor model we observed standardized rosemary extract when given orally to significantly suppress tumor growth by 46% compared to mice not receiving standardized rosemary extract. In the last several years regulatory governing bodies (e.g. European Union) have approved standardized rosemary extracts as food preservatives. These results are especially significant as it is becoming more likely that individuals will be receiving standardized rosemary extracts that are a part of a natural preservative system in various food preparations. Taken a step further, it is possible that the potential benefits that are often associated with a "Mediterranean Diet" in the future may begin to extend beyond the Mediterranean diet as more of the population is consuming standardized rosemary extracts

Selective Modulation of Endoplasmic Reticulum Stress Markers in Prostate Cancer Cells by a Standardized Mangosteen Fruit Extract

<div><p>The increased proliferation of cancer cells is directly dependent on the increased activity of the endoplasmic reticulum (ER) machinery which is responsible for protein folding, assembly, and transport. In fact, it is so critical that perturbations in the endoplasmic reticulum can lead to apoptosis. This carefully regulated organelle represents a unique target of cancer cells while sparing healthy cells. In this study, a standardized mangosteen fruit extract (MFE) was evaluated for modulating ER stress proteins in prostate cancer. Two human prostate cancer cell lines, 22Rv1 and LNCaP, and prostate epithelial cells (PrECs) procured from two patients undergoing radical prostatectomy were treated with MFE. Flow cytometry, MTT, BrdU and Western blot were used to evaluate cell apoptosis, viability, proliferation and ER stress. Next, we evaluated MFE for microsomal stability and anti-cancer activity in nude mice. MFE induced apoptosis, decreased viability and proliferation in prostate cancer cells. MFE increased the expression of ER stress proteins. Interestingly, MFE selectively promotes ER stress in prostate cancer cells while sparing PrECs. MFE suppressed tumor growth in a xenograft tumor model without obvious toxicity. Mangosteen fruit extract selectively promotes endoplasmic reticulum stress in cancer cells while sparing non-tumorigenic prostate epithelial cells. Furthermore, in an in vivo setting mangosteen fruit extract significantly reduces xenograft tumor formation.</p></div

Mangosteen fruit extract (MFE) reduced viability and inhibited proliferation <i>in vitro</i> on two different prostate cancer cell lines, 22Rv1 and LNCaP.

<p>[<b>A</b>], Microscopic pictures of MFE-treated and untreated prostate cancer cells, MFE dose was indicated. [<b>B</b>], LNCaP cells were treated with increasing doses of MFE for 24, 48, or 72 hr, cell viability was determined by MTT assay. [<b>C</b>], 22Rv1 cells were treated with increasing doses of MFE for 24, 48, or 72 hr, cell viability was determined by MTT assay. [<b>D</b>], Both 22Rv1 and LNCaP cells were treated with increasing doses of MFE, cell proliferation was evaluated by BrdU assay. Absorbance values at 450 nm (A450) represent proliferating cell numbers. These experiments were performed in triplicate and are represented by the mean along with standard deviation.</p

MFE was incubated with liver microsomes to mimic P450 Phase I metabolism in the mouse or human liver.

<p>Human and mouse liver microsomes were prepared as described in the materials and methods. Black bars represent the respective control for that individual time point. Gray bars represent the microsomal stability of α-Mangostin in Phase I enzyme system in both mouse and human liver microsomes. A ‘no preincubation’ was performed (data not shown) and did not have any impact on the interpretation of this data. [<b>A</b>], Mangosteen Fruit Extract (standardized to α-Mangostin) was incubated with mouse liver microsomes. [<b>B</b>], Mangosteen Fruit Extract (standardized to α-Mangostin) was incubated with human liver microsomes. [<b>C</b>], Twelve animals were subcutaneously injected in each flank of the mouse (i.e. two tumors per mice) with ∼1×10⁶ 22Rv1 cells to initiate tumor growth. Twenty-four hours after cell implantation, the animals in each cohort received by intraperitoneal administration vehicle or mangosteen fruit extract (35 mg/kg). Body weights of athymic nude mice treated with control vehicle or mangosteen fruit extract (35 mg/kg) intraperitoneally twice per week were measured two times weekly. [<b>D</b>], The average tumor volume of control and mangosteen fruit extract treated mice were plotted over days after tumor cell inoculation. Data points represent the mean of 12 tumors from six mice; bars represent standard deviation of the mean, *<i>P</i><0.01. [<b>E</b>], Representatives of tumor bearing mice from control and MFE-treated group.</p

Mangosteen fruit extract (MFE) induced apoptosis and apoptosis-related gene expression in prostate cancer cells.

<p>[<b>A</b>], Percentage of apoptotic cells in MFE-treated and untreated cells were determined by flow cytometry. MFE dose was 15 µg/ml. [<b>B</b>], Cleaved Caspase-3 levels in increasing doses of MFE treated 22Rv1 and LNCaP cells. Cell lysates were prepared from MFE-treated cells and cleaved caspase-3 levels were evaluated from same amounts of cell lysates by ELISA, details see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0081572#s2" target="_blank"><i>Materials and Methods</i></a>. Relative expression levels were indicated as absorbance values at 450 nm (A450). [<b>C</b>], Bax expression in increasing doses of MFE-treated 22Rv1 and LNCaP. Cell lysates were prepared from MFE-treated cells, Bax expression was detected by western blot, and β-actin was used as a loading control. These results are representatives from three independent experiments.</p