Nanoparticle delivery systems to combat drug resistance in ovarian cancer.

Due to the lack of early symptoms and difficulty of accurate diagnosis, ovarian cancer is the most lethal gynecological cancer faced by women. First-line therapy includes a combination of tumor resection surgery and chemotherapy regimen. However, treatment becomes more complex upon recurrence due to development of drug resistance. Drug resistance has been linked to many mechanisms, including efflux transporters, apoptosis dysregulation, autophagy, cancer stem cells, epigenetics, and the epithelial-mesenchymal transition. Thus, developing and choosing effective therapies is exceptionally complex. There is a need for increased specificity and efficacy in therapies for drug-resistant ovarian cancer, and research in targeted nanoparticle delivery systems aims to fulfill this challenge. Although recent research has focused on targeted nanoparticle-based therapies, few of these therapies have been clinically translated. In this review, non-viral nanoparticle delivery systems developed to overcome drug-resistance in ovarian cancer were analyzed, including their structural components, surface modifications, and drug-resistance targeted mechanisms.

Polymersomes for Therapeutic Delivery of Protein and Nucleic Acid Macromolecules: From Design to Therapeutic Applications.

Macromolecule-based therapeutic agents, particularly proteins, antigens, monoclonal antibodies, transcription factors, nucleic acids, and gene editing enzymes, have the potential to offer cures for previously untreatable diseases. However, they present an enormous delivery challenge due to poor absorption and rapid metabolism in the body. Polymersomes have tremendous potential in delivering these agents to their desired intracellular location due to increased circulation times, decreased macromolecule degradation, and decreased immune responses. In this Review, we highlight the key factors in design, development, and improved performance of these vesicles for macromolecular delivery. The recent progress made toward preclinical application of these vesicles for protein and gene delivery is also covered.

Method of limits: Female genital stretch perception thresholds.

INTRODUCTION: Women with pelvic organ prolapse describe vaginal laxity and poor sensation of vaginal tone that does not correlate with anatomical findings. This discrepancy could be explained by altered vaginal sensation and a test that could measure sensation of vaginal tone, transmitted via Aα and Aß nerve fibers, would further our understanding of the pathophysiology of vaginal laxity. OBJECTIVE: To develop quantitative sensory testing (QST) for vaginal tone using genital stretch perception thresholds (PT), assess reproducibility, and the association with age and parity. STUDY DESIGN: Prospective observational cohort study of healthy women (Canadian task force classification II-2) who underwent QST method of limits at the vagina and introitus for sensation of first awareness and stretch using a modified anorectal physiology protocol. RESULTS: Forty women underwent repeatability testing. Intra- and inter-rater repeatability using intraclass correlation coefficients (ICC) was good to excellent for both first awareness and stretch at the vagina and introitus (intra-rater ICC = 0.93, 0.95, 0.81, and 0.88, respectively; inter-rater ICC = 0.83, 0.93, 0.71, and 0.86 respectively). Normative data were collected from 100 women. Log-linear regression found a significant association between age and PT for first awareness and stretch at the vagina and introitus (P = .020, .008, .002, and <.001, respectively). There was no association with parity and PT. Nomograms were calculated using the 95% confidence limits around the regression line. CONCLUSIONS: Stretch QST is clinically feasible, valid, and reproducible. The test can be used as a tool to measure sensation in women presenting with symptoms of vaginal laxity.

Identification and characterization of a novel stay-green QTL that increases yield in maize.

Functional stay-green is a valuable trait that extends the photosynthetic period, increases source capacity and biomass and ultimately translates to higher grain yield. Selection for higher yields has increased stay-green in modern maize hybrids. Here, we report a novel QTL controlling functional stay-green that was discovered in a mapping population derived from the Illinois High Protein 1 (IHP1) and Illinois Low Protein 1 (ILP1) lines, which show very different rates of leaf senescence. This QTL was mapped to a single gene containing a NAC-domain transcription factor that we named nac7. Transgenic maize lines where nac7 was down-regulated by RNAi showed delayed senescence and increased both biomass and nitrogen accumulation in vegetative tissues, demonstrating NAC7 functions as a negative regulator of the stay-green trait. More importantly, crosses between nac7 RNAi parents and two different elite inbred testers produced hybrids with prolonged stay-green and increased grain yield by an average 0.29 megagram/hectare (4.6 bushel/acre), in 2 years of multi-environment field trials. Subsequent RNAseq experiments, one employing nac7 RNAi leaves and the other using leaf protoplasts overexpressing Nac7, revealed an important role for NAC7 in regulating genes in photosynthesis, chlorophyll degradation and protein turnover pathways that each contribute to the functional stay-green phenotype. We further determined the putative target of NAC7 and provided a logical extension for the role of NAC7 in regulating resource allocation from vegetative source to reproductive sink tissues. Collectively, our findings make a compelling case for NAC7 as a target for improving functional stay-green and yields in maize and other crops.

Enhancing drug delivery with supramolecular amphiphilic macrocycle nanoparticles: selective targeting of CDK4/6 inhibitor palbociclib to melanoma.

Drug delivery systems based on amphiphilic supramolecular macrocycles have garnered increased attention over the past two decades due to their ability to successfully formulate nanoparticles. Macrocyclic (MC) materials can self-assemble at lower concentrations without the need for surfactants and polymers, but surfactants are required to form and stabilize nanoparticles at higher concentrations. Using MCs to deliver both hydrophilic and hydrophobic guest molecules is advantageous. We developed two novel types of amphiphilic macrocycle nanoparticles (MC NPs) capable of delivering either Nile Red (NR) (a hydrophobic model) or Rhodamine B (RhB) (a hydrophilic model) fluorescent dyes. We extensively characterized the materials using various techniques to determine size, morphology, stability, hemolysis, fluorescence, loading efficiency (LE), and loading capacity (LC). We then loaded the CDK4/6 inhibitor Palbociclib (Palb) into both MC NPs using a solvent diffusion method. This yielded Palb-MC NPs in the size range of 65-90 nm. They exhibited high stability over time and in fetal bovine serum with negligible toxicity against erythrocytes. Cytotoxicity was minimal when tested against RAW macrophages, human fibroblast HDFn, and adipose stromal cells (ASCs) at higher concentrations of MC NPs. Cell viability studies were conducted with different concentrations of MC NPs, Palb-MC NPs, and free Palb against RAW macrophages, human U-87 GBM, and human M14 melanoma cell lines in vitro. Flow cytometry experiments revealed that blank MC NPs and Palb-MC NPs were selectively targeted to melanoma cells, resulting in cell death compared to the other two cell lines. Future work will focus on studying the biological effect of MC NPs including their binding affinity with molecules/receptors expressed on the M14 and other melanoma cell surfaces by molecular docking simulations. Subsequently, we will evaluate the MCs as a component of combination therapy in a murine melanoma model.

Enhancing Drug Delivery with Supramolecular Amphiphilic Macrocycle Nanoparticles: Selective Targeting of CDK4/6 Inhibitor Palbociclib to Melanoma.

Drug delivery systems based on amphiphilic supramolecular macrocycles have garnered increased attention over the past two decades due to their ability to successfully formulate nanoparticles. Macrocyclic (MC) materials can self-assemble at lower concentrations without the need for surfactants and polymers, but surfactants are required to form and stabilize nanoparticles at higher concentrations. Using MCs to deliver both hydrophilic and hydrophobic guest molecules is advantageous. We developed two novel types of amphiphilic macrocycle nanoparticles (MC NPs) capable of delivering either Nile Red (NR) (a hydrophobic model) or Rhodamine B (RhB) (a hydrophilic model) fluorescent dyes. We extensively characterized the materials using various techniques to determine size, morphology, stability, hemolysis, fluorescence, loading efficiency (LE), and loading capacity (LC). We then loaded the CDK4/6 inhibitor Palbociclib (Palb) into both MC NPs using a solvent diffusion method. This yielded Palb-MC NPs in the size range of 65-90 nm. They exhibited high stability over time and in fetal bovine serum with negligible toxicity against erythrocytes. Cytotoxicity was minimal when tested against RAW macrophages, human fibroblast HDFn , and adipose stromal cells (ASCs) at higher concentrations of MC NPs. Cell viability studies were conducted with different concentrations of MC NPs, Palb-MC NPs, and free Palb against RAW macrophages, human U-87 GBM, and human M14 melanoma cell lines in vitro. Flow cytometry experiments revealed that blank MC NPs and Palb-MC NPs were selectively targeted to melanoma cells, resulting in cell death compared to the other two cell lines. Future work will focus on studying the biological effect of MC NPs including their binding affinity with molecules/receptors expressed on the M14 and other melanoma cell surface by molecular docking simulations. Subsequently, we will evaluate the MCs as a component of combination therapy in a murine melanoma model.

Significance of an Online Evidence-Based Practice Education Module for Acute, Ambulatory, Public, and School Health Nurses.

Recent surveys of Magnet facilities and nurses found low rates of implementation of evidence-based practice in U.S. health care settings. Nursing Experts: Translating the Evidence (NExT) is a collaboration of nurses and librarians providing free online evidence-based practice nursing education benefiting nurses in all settings. The NExT online modules empowered participants to efficiently access valuable resources to inform and improve their practice in a convenient, accessible, self-paced format. Quantitative and qualitative evaluation methods and the value of collaboration are discussed. [J Contin Educ Nurs. 2023;54(4):176-184.].

Mechanisms of Drug Resistance in Ovarian Cancer and Associated Gene Targets.

In the United States, over 100,000 women are diagnosed with a gynecologic malignancy every year, with ovarian cancer being the most lethal. One of the hallmark characteristics of ovarian cancer is the development of resistance to chemotherapeutics. While the exact mechanisms of chemoresistance are poorly understood, it is known that changes at the cellular and molecular level make chemoresistance challenging to treat. Improved therapeutic options are needed to target these changes at the molecular level. Using a precision medicine approach, such as gene therapy, genes can be specifically exploited to resensitize tumors to therapeutics. This review highlights traditional and novel gene targets that can be used to develop new and improved targeted therapies, from drug efflux proteins to ovarian cancer stem cells. The review also addresses the clinical relevance and landscape of the discussed gene targets.

Peptide-based delivery of therapeutics in cancer treatment.

Current delivery strategies for cancer therapeutics commonly cause significant systemic side effects due to required high doses of therapeutic, inefficient cellular uptake of drug, and poor cell selectivity. Peptide-based delivery systems have shown the ability to alleviate these issues and can significantly enhance therapeutic loading, delivery, and cancer targetability. Peptide systems can be tailor-made for specific cancer applications. This review describes three peptide classes, targeting, cell penetrating, and fusogenic peptides, as stand-alone nanoparticle systems, conjugations to nanoparticle systems, or as the therapeutic modality. Peptide nanoparticle design, characteristics, and applications are discussed as well as peptide applications in the clinical space.

Fusogenic peptide delivery of bioactive siRNAs targeting CSNK2A1 for treatment of ovarian cancer.

Ovarian cancer has shown little improvement in survival among advanced-stage patients over the past decade. Current treatment strategies have been largely unsuccessful in treating advanced disease, with many patients experiencing systemic toxicity and drug-resistant metastatic cancer. This study evaluates novel fusogenic peptide carriers delivering short interfering RNA (siRNA) targeting casein kinase II, CSNK2A1, for reducing the aggressiveness of ovarian cancer. The peptides were designed to address two significant barriers to siRNA delivery: insufficient cellular uptake and endosomal entrapment. The three peptide variants developed, DIVA3, DIV3H, and DIV3W, were able to form monodisperse nanoparticle complexes with siRNA and protect siRNAs from serum and RNase degradation. Furthermore, DIV3W demonstrated optimal delivery of bioactive siRNAs into ovarian cancer cells with high cellular uptake efficiency and mediated up to 94% knockdown of CSNK2A1 mRNA compared with non-targeting siRNAs, resulting in decreased cell migration and recolonization in vitro. Intratumoral delivery of DIV3W-siCSNK2A1 complexes to subcutaneous ovarian tumors resulted in reduced CSNK2A1 mRNA and CK2α protein expression after 48 h and reduced tumor growth and migration in a 2-week multi-dosing regimen. These results demonstrate the potential of the DIV3W peptide to deliver bioactive siRNAs and confirms the role of CSNK2A1 in cell-cell communication and proliferation in ovarian cancer.

Factors Affecting Secondary and Supramolecular Structures of Self-Assembling Peptide Nanocarriers.

Self-assembling peptides are a popular vector for therapeutic cargo delivery due to their versatility, tunability, and biocompatibility. Accurately predicting secondary and supramolecular structures of self-assembling peptides is essential for de novo peptide design. However, computational modeling of such assemblies is not yet able to accurately predict structure formation for many peptide sequences. This review identifies patterns in literature between secondary and supramolecular structures, primary sequences, and applications to provide a guide for informed peptide design. An overview of peptide structures, their applications as nanocarriers, and analytical methods for characterizing secondary and supramolecular structure is examined. A top-down approach is then used to identify trends between peptide sequence and assembly structure from the current literature, including an analysis of the drivers at work, such as local and nonlocal sequence effects and solution conditions.

De Novo Self-Assembling Peptides Mediate the Conversion of Temozolomide and Delivery of a Model Drug into Glioblastoma Multiforme Cells.

Glioblastoma multiforme (GBM) is the most aggressive central nervous system tumor, and standard treatment, including surgical resection, radiation, and chemotherapy, has not significantly improved patient outcomes over the last 20 years. Temozolomide (TMZ), the prodrug most commonly used to treat GBM, must pass the blood-brain barrier and requires a basic pH to convert to its active form. Due to these barriers, less than 30% of orally delivered TMZ reaches the central nervous system and becomes bioactive. In this work, we have developed a novel biomaterial delivery system to convert TMZ to its active form and that shows promise for intracellular TMZ delivery. Self-assembling peptides were characterized under several different assembly conditions and evaluated for TMZ loading and conversion. Both solvent and method of assembly were found to affect the supramolecular and secondary structure of peptide assemblies. Additionally, as peptides degraded in phosphate-buffered saline, TMZ was rapidly converted to its active form. This work demonstrates that peptide-based drug delivery systems can effectively create a local stimulus during drug delivery while remaining biocompatible. This principle could be used in many future biomedical applications in addition to cancer treatment, such as wound healing and regenerative medicine.

Parental perception on the efficacy of a physical activity program for preschoolers.

Childhood obesity is among the leading health concerns in the United States. The relationship between unmet physical activity needs in young children is of particular interest as the trend in childhood obesity continues to rise and unmet physical activity needs are identified. The preschool years are an influential time in promoting healthful lifestyle habits and early childhood interventions may help establish lifelong healthful behaviors which could help prevent obesity later in life. The Food Friends®: Get Movin' with Mighty Moves® is a preschool physical activity program which aims to improve children's gross motor skills and physical activity levels. The home environment and parental modeling are critical factors related to child physical activity in this population. The parent component, Mighty Moves®: Fun Ways to Keep Families Active and Healthy, was designed to address barriers in the home environment that lead to unmet physical activity needs in preschoolers and their families. The program and materials were designed based on Social Marketing tenets and Social Learning Theory principles. Four Colorado Head Start centers were assigned to an experimental group as part of the Mighty Moves® group randomized trial. Quantitative and qualitative evaluation methods were used to determine what messages and materials reached and motivated the target audience to increase physical activity levels. Results of the study indicated the program's materials helped families and children to be more physically active. Additionally, materials and material dissemination were revised to enhance program goals.

In Situ Photopolymerization of Acrylamide Hydrogel to Coat Cellulose Acetate Nanofibers for Drug Delivery System.

In this study we developed electrospun cellulose acetate nanofibers (CANFs) that were loaded with a model non-steroidal anti-inflammatory drug (NSAID) (ibuprofen, Ib) and coated with poly(acrylamide) (poly-AAm) hydrogel polymer using two consecutive steps: an electrospinning process followed by photopolymerization of AAm. Coated and non-coated CANF formulations were characterized by several microscopic and spectroscopic techniques to evaluate their physicochemical properties. An analysis of the kinetic release profile of Ib showed noticeable differences due to the presence or absence of the poly-AAm hydrogel polymer. Poly-AAm coating facilitated a constant release rate of drug as opposed to a more conventional burst release. The non-coated CANFs showed low cumulative drug release concentrations (ca. 35 and 83% at 5 and 10% loading, respectively). Conversely, poly-AAm coated CANFs were found to promote the release of drug (ca. 84 and 99.8% at 5 and 10% loading, respectively). Finally, the CANFs were found to be superbly cytocompatible.

Classifying changes in LN-18 glial cell morphology: a supervised machine learning approach to analyzing cell microscopy data via FIJI and WEKA.

In cell-based research, the process of visually monitoring cells generates large image datasets that need to be evaluated for quantifiable information in order to track the effectiveness of treatments in vitro. With the traditional, end-point assay-based approach being error-prone, and existing computational approaches being complex, we tested existing machine learning frameworks to find methods that are relatively simple, yet powerful enough to accomplish the goal of analyzing cell microscopy data. This paper details the machine learning pipeline for pixel-based classification and object-based classification. Furthermore, it compares the performances of three classifiers. The classifiers evaluated were the fast-random forest (RF), the sequential minimal optimization (SMO), and the Bayesian network (BN). Images were first preprocessed using smoothing and contrast methods found in FIJI. For pixel-based classification, the preprocessed images were fed into the Trainable Waikato Segmentation (TWS). For object-based classification, training and classification were conducted within the Waikato Environment for Knowledge Analysis (WEKA) interface. All classifiers' performance was evaluated using the WEKA experimental explorer. In terms of performance, the BN had the lowest classification accuracy for both the pixel-based and object-based model. The object-based SMO classifier had the best performance with the lowest mean absolute error of 0.05. The TWS and WEKA interface allows users to easily create and train classifiers for image analysis. However, for analyzing large image datasets, they are not ideal. Grapical abstract.

Vectors for Glioblastoma Gene Therapy: Viral & Non-Viral Delivery Strategies.

Glioblastoma multiforme is the most common and aggressive primary brain tumor. Even with aggressive treatment including surgical resection, radiation, and chemotherapy, patient outcomes remain poor, with five-year survival rates at only 10%. Barriers to treatment include inefficient drug delivery across the blood brain barrier and development of drug resistance. Because gliomas occur due to sequential acquisition of genetic alterations, gene therapy represents a promising alternative to overcome limitations of conventional therapy. Gene or nucleic acid carriers must be used to deliver these therapies successfully into tumor tissue and have been extensively studied. Viral vectors have been evaluated in clinical trials for glioblastoma gene therapy but have not achieved FDA approval due to issues with viral delivery, inefficient tumor penetration, and limited efficacy. Non-viral vectors have been explored for delivery of glioma gene therapy and have shown promise as gene vectors for glioma treatment in preclinical studies and a few non-polymeric vectors have entered clinical trials. In this review, delivery systems including viral, non-polymeric, and polymeric vectors that have been used in glioblastoma multiforme (GBM) gene therapy are discussed. Additionally, advances in glioblastoma gene therapy using viral and non-polymeric vectors in clinical trials and emerging polymeric vectors for glioma gene therapy are discussed.

Dual peptide-mediated targeted delivery of bioactive siRNAs to oral cancer cells in vivo.

OBJECTIVES: Despite significant advances in cancer treatment, the prognosis for oral cancer remains poor in comparison to other cancer types, including breast, skin, and prostate. As a result, more effective therapeutic modalities are needed for the treatment of oral cancer. Consequently, in the present study, we examined the feasibility of using a dual peptide carrier approach, combining an epidermal growth factor receptor (EGFR)-targeting peptide with an endosome-disruptive peptide, to mediate targeted delivery of small interfering RNAs (siRNAs) into EGFR-overexpressing oral cancer cells and induce silencing of the targeted oncogene, cancerous inhibitor of protein phosphatase 2A (CIP2A). MATERIALS AND METHODS: Fluorescence microscopy, real-time PCR, Western blot analysis, and in vivo bioimaging of mice containing orthotopic xenograft tumors were used to examine the ability of the dual peptide carrier to mediate specific delivery of bioactive siRNAs into EGFR-overexpressing oral cancer cells/tissues. RESULTS: Co-complexation of the EGFR-targeting peptide, GE11R9, with the endosome-disruptive 599 peptide facilitated the specific uptake of siRNAs into oral cancer cells overexpressing EGFR in vitro with optimal gene silencing observed at a 60:30:1 (GE11R9:599:siRNA) molar ratio. Furthermore, when administered systemically to mice bearing xenograft oral tumors, this dual peptide complex mediated increased targeted delivery of siRNAs into tumor tissues in comparison to the 599 peptide alone and significantly enhanced CIP2A silencing. CONCLUSION: Herein we provide the first report demonstrating the clinical potential of a dual peptide strategy for siRNA-based therapeutics by synergistically mediating the effective targeting and delivery of bioactive siRNAs into EGFR-overexpressing oral cancer cells.

Building Capacity in Community-Based Participatory Research Partnerships Through a Focus on Process and Multiculturalism.

BACKGROUND: In health research, investigators and funders are emphasizing the importance of collaboration between communities and academic institutions to achieve health equity. Although the principles underlying community-academic partnered research have been well-articulated, the processes by which partnerships integrate these principles when working across cultural differences are not as well described. OBJECTIVES: We present how Project GRACE (Growing, Reaching, Advocating for Change and Empowerment) integrated participatory research principles with the process of building individual and partnership capacity. METHODS: We worked with Vigorous Interventions In Ongoing Natural Settings (VISIONS) Inc., a process consultant and training organization, to develop a capacity building model. We present the conceptual framework and multicultural process of change (MPOC) that was used to build individual and partnership capacity to address health disparities. CONCLUSIONS: The process and capacity building model provides a common language, approach, and toolset to understand differences and the dynamics of inequity. These tools can be used by other partnerships in the conduct of research to achieve health equity.

Fusogenic-oligoarginine peptide-mediated silencing of the CIP2A oncogene suppresses oral cancer tumor growth in vivo.

Intracellular delivery and endosomal escape of functional small interfering RNAs (siRNAs) remain major barriers limiting the clinical translation of RNA interference (RNAi)-based therapeutics. Recently, we demonstrated that a cell-penetrating endosome-disruptive peptide we synthesized, termed 599, enhanced the intracellular delivery and bioavailability of siRNAs designed to target the CIP2A oncoprotein (siCIP2A) into oral cancer cells and consequently inhibited oral cancer cell invasiveness and anchorage-independent growth in vitro. Thus, to further assess the therapeutic potential of the 599 peptide in mediating RNAi-based therapeutics for oral cancer and its prospective applicability in clinical settings, the objective of the current study was to determine whether intratumoral dosing of the 599 peptide-siCIP2A complex could induce silencing of CIP2A and consequently impair tumor growth using a xenograft oral cancer mouse model. Our results demonstrate that the 599 peptide is able to protect siRNAs from degradation by serum and ribonucleases in vitro and upon intratumoral injection in vivo, confirming the stability of the 599 peptide-siRNA complex and its potential for therapeutic utility. Moreover, 599 peptide-mediated delivery of siCIP2A to tumor tissue induces CIP2A silencing without any associated toxicity, consequently resulting in reduction of the mitotic index and significant inhibition of tumor growth. Together, these data suggest that the 599 peptide carrier is a clinically effective mediator of RNAi-based cancer therapeutics.

Bioengineering strategies for designing targeted cancer therapies.

The goals of bioengineering strategies for targeted cancer therapies are (1) to deliver a high dose of an anticancer drug directly to a cancer tumor, (2) to enhance drug uptake by malignant cells, and (3) to minimize drug uptake by nonmalignant cells. Effective cancer-targeting therapies will require both passive- and active-targeting strategies and a thorough understanding of physiologic barriers to targeted drug delivery. Designing a targeted therapy includes the selection and optimization of a nanoparticle delivery vehicle for passive accumulation in tumors, a targeting moiety for active receptor-mediated uptake, and stimuli-responsive polymers for control of drug release. The future direction of cancer targeting is a combinatorial approach, in which targeting therapies are designed to use multiple-targeting strategies. The combinatorial approach will enable combination therapy for delivery of multiple drugs and dual ligand targeting to improve targeting specificity. Targeted cancer treatments in development and the new combinatorial approaches show promise for improving targeted anticancer drug delivery and improving treatment outcomes.