Renee Coffman

Report of the 2010-2011 Standing Committee on Advocacy: Leveraging faculty engagement to improve public policy.

According to the Bylaws of the American Association of Colleges of Pharmacy (AACP), the Advocacy Committee: “will advise the Board of Directors on the formation of positions on matters of public policy and on strategies to advance those positions to the public and private sectors on behalf of academic pharmacy.” COMMITTEE CHARGE President Rodney Carter charged the 2010-2011 Advocacy Committee to: “examine the question how can AACP and its members most effectively leverage faculty scholarship/research to impact on public policy at the state and federal level?” The Committee met in-person in October in Arlington, VA to discuss the charge and determine the approach to meeting the charge. After a wide-ranging discussion guided by the Chair the Committee agreed that a case study approach would meet the intent of the charge and serve the broader Academy by providing examples of evidence-based advocacy. Committee members agreed that the case studies could include completed, ongoing, or developing examples of how faculty scholarship and research did or failed to impact public policy. A framework for case study submission was developed and agreed to by the Committee. Case Study Framework: Each advocacy committee member will present one initiative that supports the integration of the pharmacist or recognizes academic pharmacy as a resource for evidence-based public policy development as a case study that provides a “roadmap for implementation” for AACP members. Each case study will be included as a section in the report. Each section will use the following format: State the healthcare reform/advocacy issue and the opportunity or expectation for the integration of the pharmacist; Describe the development of the partnership with the academic or community-based partner and their understanding and expectation of the integration of the pharmacist into issue activities or how pharmacy faculty can contribute to furthering public policy development; Describe through examples of teaching, research, or service, current activity at the college or school level to address the issue; List the AACP/other resources that provide evidence of academic activity that support the selected issue; and Recommendations regarding additional resources or evidence needed to advance the role of the pharmacist into the activities supporting the healthcare reform/advocacy initiative. BACKGROUND Leveraging public policy development to your advantage requires strong evidence that supports or opposes the policy. Public policy is advanced by science-based contributions.1,2 It is helpful to keep in mind that there is no guarantee that evidence improves the final policy since politics can be a dominant influence. However the strength of the evidence can contribute to its consideration in public policy.3 How and to whom the evidence is presented remains an essential element of influencing public policy. The creation of new knowledge and evaluation of existing knowledge are responsibilities of every faculty member of a college or school of pharmacy. Therefore, leveraging public policy development requires 1) identification of public policy of personal or professional interest and those supporting or opposing the policy, 2) assessing the policy for personal or collective contribution opportunities, and 3) determining the best approach for contributing the evidence. For instance, a significant piece of public policy, the Patient Protection and Affordable Care Act, includes provisions that seek to increase access to medication therapy management. The specific public concern being poor medication management is costly in terms of health and economic outcomes. This public concern was leveraged by evidence generated, translated and provided by pharmacy faculty. This evidence included examples of research, some of it supported by federal grants,4 demonstrating improved health outcomes associated with the provision of MTM services. Influencing public policy through evidence-sharing will continue to be an important goal of academic and professional organizations. The Patient Protection and Affordable Care Act creates many opportunities for academic pharmacy to leverage its implementation through the creation of new knowledge or evaluate current knowledge and translating both new and current knowledge into programs and services that meet the intent of the law.5 Members of the Academy are already providing significant contributions to the literature supporting the integration of the pharmacist across the continuum of care competent to provide patient-centered, team-based care.6,7 Likewise, daily activities such as interactions with state-based organizations, community partners, health insurance payers and even accrediting organizations provide opportunities for your position to be articulated and supported through evidence-sharing. The ability to leverage policy development requires an understanding and appreciation of other individuals and groups that will be engaged in the creation of new policy and its eventual implementation. Understanding and appreciation, regardless of whether you agree or disagree, requires the development of relationships with individuals, institutions and organizations involved in influencing public policy you deem important or relevant to your personal or professional goals. Understanding the advocacy or public policy goals of others is the first step in determining how what information you will provide to leverage their goals to your advantage. Through discussions with other individuals and groups you begin the second step, to assess the relevance of their goals to yours. Identification of goal alignment is an important step toward influencing public policy. Shared goals strengthen advocacy. The recognition of this strength regularly results in the establishment of coalitions and task forces that combine individual or organizational goals into a larger presence to influence public policy development, implementation and evaluation. Coalitions and other groups built upon mutual advocacy goals are sustained and strengthened through the evidence they are able to share with those developing or implementing the policy. Toward this end, pharmacy faculty are able to leverage public policy by working with others that share their goals, creating and communicating evidence that supports those goals.

Student Stress in a Three-Year Doctor of Pharmacy Program Using a Mastery Learning Educational Model

Objective. To identify stress and stress-relieving mechanisms among second-year pharmacy students in a 3-year doctor of pharmacy (PharmD) program using a Mastery Learning Educational Model and to compare findings with those from a 4-year program. Methods. Second-year PharmD students in a 3-year program were asked to complete a series of questionnaires including the Perceived Stress Scale (PSS) regarding stress and stress-relieving activities. Results. The average PSS score for the 3-year PharmD cohort was significantly higher than the score of demographically similar students enrolled in a 4-year PharmD program (P = 0.04). There were significant differences between the 2 groups’ scores on 5 items on the PSS including how often they: were upset because something happened unexpectedly, felt unable to control important things, felt nervous and stressed, thought about things that had to be accomplished, and were able to control the way they spent their time. The rate of prescription drug misuse among those in the 3-year PharmD program was 11.6%. Conclusions. Students in a 3-year PharmD program with a unique educational model experienced more stress than students in a traditional 4-year PharmD program.

Abstract B01: Phosphorylation of pro-oncogenic c-Src at serine-17 by protein kinase G-Iα promotes chemoresistance/cell proliferation in human non-small cell lung cancer cells.

We previously showed that PKG-Iα promotes cell survival in neural cells (N1E-115 neuroblastoma and NG108-15 neuroblastoma-glioma hybrid cells) and significantly contributes to serine-155 phosphorylation of BAD, an apoptosis-regulating protein (Johlfs and Fiscus 2010). We also found, by using both gene knockdown and pharmacological inhibitors (ODQ, DT-2 and DT-3) to decrease PKG-Iα kinase activity, that PKG-Iα promotes cell proliferation and chemoresistance in human ovarian cancer cells, involving a novel interaction between PKG-Iα and the oncogenic protein c-Src (Fiscus et al 2012, Leung et al 2010). PKG-Iα activity was dependent on c-Src-catalyzed tyrosine phosphorylation of PKG-Iα; and, reciprocally, c-Src activation was dependent on expression and kinase activity of PKG-Iα;. We hypothesized that activation of c-Src involves PKG-Iα-catalyzed phosphorylation at serine-17 of c-Src, because amino acids around serine-17 provide consensus sequence for PKG-I, and tαhat this plays an important role in proliferation and chemoresistance. As c-Src has been proposed to promote cell proliferation and chemoresistance in NSCLC cells, the present study aimed to: 1) determine if c-Src increases PKG-Iα kinase activity in NSCLC cells using our newly-developed near-infrared-fluorescence (NIRF)-kinase assay, 2) determine if serine-17-phosphorylation of c-Src, in intact cancer cells, is dependent on PKG-Iα expression and kinase activity (using siRNA gene knockdown and pharmacological inhibitors), and 3) determine if inhibition/knockdown of c-Src and PKG-Iα alters chemoresistance and proliferation of NSCLC cells. Protein expression of c-Src and PKG-Iα was determined by conventional Western blot analysis and new ultrasensitive capillary-electrophoresis-based NanoPro100/1000 (ProteinSimple, Santa Clara, CA, USA). NanoPro100/1000 allows clear separation and identification of different c-Src and PKG-Iαphospho-forms (a novel way of identifying the activation of c-Src and PKG-Iα). We found PKG-Iα was able to directly catalyze phosphorylation of serine-17 of c-Src, increasing autophosphorylation of c-Src at tyrosine-419, the activation site of c-Src. Inhibition of PKG-Iα kinase activity using DT-2 or silencing of PKG-Iα expression using siRNA dramatically reduced the intracellular phosphorylation of c-Src at serine-17. Both kinase-activity inhibition and knockdown of PKG-Iα caused significant increases in apoptosis, synergistically enhancing apoptosis induced by the chemotherapeutic agent cisplatin, and dramatically decreased cell proliferation/colony formation in NSCLC cells. This novel reciprocal relationship between c-Src and PKG-Iα provides a new target for the development of new anti-cancer therapeutic agents. Fiscus RR, Leung EL, Wong JC, Johlfs MG (2012). Nitric oxide/protein kinase G-Ialpha promote c-Src activation, proliferation and chemoresistance in ovarian cancer. In: Farghaly S (ed). Ovarian Cancer - Basic Science Perspective. Intech Open Access Publisher. pp 315-334. Johlfs MG, Fiscus RR (2010). Protein kinase G type-Ialpha phosphorylates the apoptosis-regulating protein Bad at serine 155 and protects against apoptosis in N1E-115 cells. Neurochem Int 56: 546-553. Leung EL, Wong JC, Johlfs MG, Tsang BK, Fiscus RR (2010). Protein kinase G type Ialpha activity in human ovarian cancer cells significantly contributes to enhanced Src activation and DNA synthesis/cell proliferation. Mol Cancer Res 8: 578-591. Citation Format: Mary G. Johlfs, Ronald R. Fiscus, Priyatham Gorjala, Renee Coffman, Harry Rosenberg. Phosphorylation of pro-oncogenic c-Src at serine-17 by protein kinase G-Iα promotes chemoresistance/cell proliferation in human non-small cell lung cancer cells. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr B01.

Abstract A29: Advanced nano-proteomics based on ultrasensitive CE toquantitate protein expression/phosphorylation levels: Discovery of novel proteins involved in chemoresistance/invasion of lung cancer cells.

Our laboratory has shown that resistance to chemotherapeutic agent cisplatin (chemoresistance) in non-small cell lung cancer (NSCLC) cells involves exaggerated phosphorylation of transcription factor CREB and high-level expression of pro-survival/anti-apoptotic proteins c-IAP1, livin, Mcl-1 and survivin, resulting from hyperactivation of a novel serine/threonine kinase, protein kinase G-Iα (PKG-Iα) (Wong et al. 2012). PKG-Iα also promotes cell proliferation/DNA synthesis and migration/invasion in both NSCLC and ovarian cancer cells (Fiscus et al., 2012; Wong et al. 2012). Because of low expression levels, these proteins are difficult to accurately quantify by conventional Western blotting. To solve this problem, we are developing new applications for NanoPro 100 and NanoPro 1000 (ProteinSimple), state-of-the-art instruments based on advanced “nano-proteomics” ultilizing ultrasensitive capillary electrophoresis (CE)-based technologies (Fiscus and Johlfs, 2012; Fiscus et al., 2012). The NanoPro 100/NanoPro 1000 are capable of accurately quantifying protein expression/phosphorylation levels with sensitivities >100-times and >500-times, respectively, that of conventional Western blots, thus allowing, for the first time, accurate measurements of lower-abundance proteins (e.g. PKG-Iα, survivin and other pro-survival proteins). Extremely small samples of Wong JC, Bathina M and RR Fiscus (2012) Cyclic GMP/protein kinase G type-Iα (PKG-Iα signaling pathway promotes CREB phosphorylation and maintains higher c-IAP1, livin, survivin and Mcl-1 expression and the inhibition of PKG-Iα kinase activity synergizes with cisplatin in non-small cell lung cancer cells. J Cell Biochem. 113: 3587-3598. Fiscus RR, Leung EL, Wong JC and MG Johlfs (2012) Nitric Oxide/Protein Kinase G-IαPromotes c-Src Activation, Proliferation and Chemoresistance in Ovarian Cancer. In: Farghaly, S. (ed). Ovarian Cancer - Basic Science Perspective. Intech Open Access Publisher. pp 315-334. Fiscus RR and MG Johlfs (2012) Protein Kinase G (PKG): Involvement in Promoting Neural Cell Survival, Proliferation, Synaptogenesis and Synaptic Plasticity and the Use of New Ultrasensitive Capillary-Electrophoresis-based Methodologies for Measuring PKG Expression and Molecular Actions. In: Mukai H (ed). Protein Kinase Technologies, in NEUROMETHODS series, Springer. pp 319-348. Citation Format: Ronald R. Fiscus, Mary G. Johlfs, Janica C. Wong, Renee Coffman, Harry Rosenberg. Advanced nano-proteomics based on ultrasensitive CE toquantitate protein expression/phosphorylation levels: Discovery of novel proteins involved in chemoresistance/invasion of lung cancer cells. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr A29.

Abstract 4558: Resveratrol alters the kinase activity of PKG-Iα and Src family in A2780cp cells (ovarian cancer cell line with mutated p53) resulting in growth inhibition

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Epithelial ovarian cancer is a leading cause of death in gynecological cancer patients. One major hurdle in treating ovarian cancer is drug resistance to commonly-used chemotherapy like cisplatin. Resveratrol (3,5,4′-trihydroxy-trans-stilbene), a natural polyphenol found in foods/drinks like grapes, berries, peanuts and red wine, can inhibit tumor growth. Our lab has previously shown that resveratrol down-regulates expression of protein kinase G type-I (PKG-I) in cancer cells, which correlated with decreased cell proliferation and increased apoptosis. For the present study, we used A2780cp cells, which are cisplatin-resistant epithelial ovarian cancer cells with mutated p53. Our data show that resveratrol did not induce apoptosis even at a higher concentration of 500 µM. However, resveratrol caused concentration-dependent decreases (down to 50% of control levels at 100 µM) of both DNA synthesis and cell adhesion. We also observed a concentration-dependent increase in protein expression levels of PKG-I to 3 times of control levels at 50 µM resveratrol, but a drop at 100 µM resveratrol. Though total Src remained fairly unchanged, pSrcY416 (phosphorylation at tyrosine-416) and pSrcS17 (phosphorylation at serine-17), indicators of Src activation, decreased at 50 and 100 µM resveratrol [assessed by ultrasensitive quantitative NanoPro 1000 (ProteinSimple), a new state-of-the-art capillary electrophoresis-based protein analysis system]. Decreased pSrcY416 levels (44% of control levels at 100 µM resveratrol) suggest that the kinase activity of Src family kinases are decreased at the higher concentrations of resveratrol that cause inhibition of ovarian cancer cell proliferation. These results suggest that resveratrol can be effective in decreasing cell proliferation in chemoresistant ovarian cancer cells and that the mechanism of action of resveratrol may involve a down-regulation of the protein expression levels of PKG-I and the typrosine-kinase activity of Src family of kinases. Citation Format: Priyatham Gorjala, Janica C. Wong, Benjamin F.b Constantino, Mary G. Johlfs, Renee Coffman, Harry Rosenberg, Ronald Fiscus. Resveratrol alters the kinase activity of PKG-Iα and Src family in A2780cp cells (ovarian cancer cell line with mutated p53) resulting in growth inhibition. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4558. doi:10.1158/1538-7445.AM2014-4558

Abstract 1008: Resveratrol causes biphasic apoptotic and proliferative effects and at higher/proapoptotic/anti-angiogenesis concentrations causes suppression of nitric oxide/cGMP/protein kinase G signaling and decreased expression of prosurvival proteins c-IAP1, c-IAP2, livin and XIAP in human umbilical vein endothelial cells

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Anti-cancer/anti-angiogenesis effects of resveratrol have been reported. Resveratrol increases nitric oxide (NO) production via increased expression of endothelial-form-NO-synthase (eNOS). However, the role of cGMP/protein kinase G (PKG) signaling, a pathway activated by NO/eNOS, in the anti-angiogenic effects of resveratrol is still unclear. In the present study, involvement of endogenous NO/cGMP/PKG pathway and downstream pro-survival proteins (Inhibitor of Apoptosis Proteins, IAPs) are studied in relation to anti-angiogenic effects of resveratrol in human umbilical vein endothelial cells (HUVECs). Our results show that resveratrol at higher/anti-angiogenesis concentrations (>50 μM) inhibits tube formation and cell migration/invasion. Resveratrol stimulates proliferation/DNA synthesis at lower concentration (10 µM), but inhibits proliferation/DNA synthesis at higher concentrations (100 and 500 µM). Moreover, resveratrol at lower concentrations (10, 20 and 50 µM) protects HUVECs against spontaneous apoptosis, but induces higher-level apoptosis at higher concentration (500 µM). ODQ (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one), inhibitor of endogenous NO-induced activation of soluble-guanylyl-cyclase and subsequent cGMP biosynthesis, completely abolishes the anti-apoptotic/cytoprotective effects of low-concentration resveratrol. 8-Br-cGMP, a direct stimulator of PKG kinase-activity, protects against pro-apoptotic effects of high-concentration resveratrol, establishing a cytoprotective role of activated-PKG in HUVECs. Western blot analyses show that higher/pro-apoptotic/anti-angiogenesis concentrations of resveratrol cause suppression of PKG kinase-activity (indicated by decreased VASP phosphorylation at Ser239) and decreased expression of eNOS and four IAPs, c-IAP1, c-IAP2, livin and XIAP. Overall, our results suggest a cytoprotective role of NO/cGMP/PKG signaling in HUVECs and further suggest that resveratrol (>50 μM)-induced anti-angiogenesis/pro-apoptosis involves suppression of NO/cGMP/PKG signaling and decreased expression of pro-survival proteins c-IAP1, c-IAP2, livin and XIAP. Citation Format: Janica C. Wong, Renee Coffman, Harry Rosenberg, Ronald R. Fiscus. Resveratrol causes biphasic apoptotic and proliferative effects and at higher/proapoptotic/anti-angiogenesis concentrations causes suppression of nitric oxide/cGMP/protein kinase G signaling and decreased expression of prosurvival proteins c-IAP1, c-IAP2, livin and XIAP in human umbilical vein endothelial cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1008. doi:10.1158/1538-7445.AM2014-1008