Needle-Free Dermal Delivery of a Diphtheria Toxin CRM197Mutant on Potassium-Doped Hydroxyapatite Microparticles

ABSTRACTInjections with a hypodermic needle and syringe (HNS) are the current standard of care globally, but the use of needles is not without limitation. While a plethora of needle-free injection devices exist, vaccine reformulation is costly and presents a barrier to their widespread clinical application. To provide a simple, needle-free, and broad-spectrum protein antigen delivery platform, we developed novel potassium-doped hydroxyapatite (K-Hap) microparticles with improved protein loading capabilities that can provide sustained local antigen presentation and release. K-Hap showed increased protein adsorption over regular hydroxyapatite (P< 0.001), good structural retention of the model antigen (CRM197) with 1% decrease in α-helix content and no change in β-sheet content upon adsorption, and sustained releasein vitro. Needle-free intradermal powder inoculation with K-Hap–CRM197induced significantly higher IgG1 geometric mean titers (GMTs) than IgG2a GMTs in a BALB/c mouse model (P< 0.001) and induced IgG titer levels that were not different from the current clinical standard (P> 0.05), namely, alum-adsorbed CRM197by intramuscular (i.m.) delivery. The presented results suggest that K-Hap microparticles may be used as a novel needle-free delivery vehicle for some protein antigens.

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Antitumor Activity of Nanoparticles Loaded with PHT-427, a Novel AKT/PDK1 Inhibitor, for the Treatment of Head and Neck Squamous Cell Carcinoma

Pharmaceutics ◽

10.3390/pharmaceutics13081242 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1242

Author(s):

Joaquín Yanes-Díaz ◽

Raquel Palao-Suay ◽

María Rosa Aguilar ◽

Juan Ignacio Riestra-Ayora ◽

Antonio Ferruelo-Alonso ◽

...

Keyword(s):

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Head And Neck ◽

Squamous Cell ◽

Polymeric Nanoparticles ◽

Growth Factor Receptor ◽

Standard Of Care ◽

Neck Squamous Cell Carcinoma ◽

Current Standard

Currently, new treatments are required to supplement the current standard of care for head and neck squamous cell carcinoma (HNSCC). The phosphatidylinositol3-kinase (PI3K) signaling pathway is commonly altered and activated in HNSCC. PHT-427 is a dual PI3K-mammalian target of the AKT/PDK1 inhibitor; however, to the best of our knowledge, the effect of the PHT-427 inhibitor on HNSCC has not been investigated. This study aims to evaluate the antitumoral effect of PHT-427-loaded polymeric nanoparticles based on α-tocopheryl succinate (α-TOS). The in vitro activity of PHT-427 was tested in hypopharynx carcinoma squamous cells (FaDu) to measure the cell viability, PI3KCA/AKT/PDK1 gene expression, and PI3KCA/AKT/PDK1 levels. Apoptosis, epidermal growth factor receptor (EGFR), and reactive oxygen species (ROS) were also measured. The presence of PHT-427 significantly enhances its antiproliferative and proapoptotic activity by inactivating the PI3K/AKT/PDK1 pathway. Nanoparticles (NPs) effectively suppress AKT/PDK1 expression. Additionally, NPs loaded with PHT-427 produce high oxidative stress levels that induce apoptosis. In conclusion, these results are promising in the use of this nanoformulation as a PHT-427 delivery system for effective HNSCC treatment.

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Novel Neonatal Umbilical Catheter Protection and Stabilization Device in In vitro Model of Catheterized Human Umbilical Cords: Effect of Material and Venting on Bacterial Colonization

American Journal of Perinatology ◽

10.1055/s-0039-1700542 ◽

2019 ◽

Author(s):

Lauren S. Y. Wood ◽

Janene H. Fuerch ◽

Carl L. Dambkowski ◽

Eric F. Chehab ◽

Shivani Torres ◽

...

Keyword(s):

Bacterial Colonization ◽

Bacterial Load ◽

Standard Of Care ◽

Adhesive Tape ◽

Human Umbilical Cord ◽

Current Standard ◽

Central Lines ◽

Umbilical Cords

Abstract Objective Umbilical central lines deliver life-saving medications and nutrition for neonates; however, complications associated with umbilical catheters (UCs) occur more frequently than in adults with central lines (i.e., line migration, systemic infection). We have developed a device for neonatal UC protection and stabilization to reduce catheter exposure to bacteria compared with the standard of care: “goal post” tape configuration. This study analyzes the effect of device venting and material on bacterial load of human umbilical cords in vitro. Study Design Catheters were inserted into human umbilical cord segments in vitro, secured with plastic or silicone vented prototype versus tape, and levels of bacterial colonization were compared between groups after 7 days of incubation. Results Nonvented plastic prototype showed increased bacterial load compared with goal post (p = 0.04). Colonization was comparable between the goal post and all vented plastic prototypes (p ≥ 0.30) and when compared with the vented silicone device (p = 1). Conclusion A novel silicone device does not increase external bacterial colonization compared with the current standard of care for line securement, and may provide a safe, convenient alternative to standard adhesive tape for UC stabilization. Future studies are anticipated to establish safety in vivo, alongside benefits such as migration and infection reduction.

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CD99 Expression in Glioblastoma Molecular Subtypes and Role in Migration and Invasion

International Journal of Molecular Sciences ◽

10.3390/ijms20051137 ◽

2019 ◽

Vol 20 (5) ◽

pp. 1137 ◽

Cited By ~ 3

Author(s):

Lais Cardoso ◽

Roseli Soares ◽

Talita Laurentino ◽

Antonio Lerario ◽

Suely Marie ◽

...

Keyword(s):

Focal Adhesion ◽

Molecular Subtypes ◽

Transmembrane Protein ◽

Standard Of Care ◽

Actin Dynamics ◽

In Vitro Assays ◽

Migration And Invasion ◽

Current Standard ◽

Tumor Cell Migration

Glioblastoma (GBM) is the most aggressive type of brain tumor, with an overall survival of 17 months under the current standard of care therapy. CD99, an over-expressed transmembrane protein in several malignancies, has been considered a potential target for immunotherapy. To further understand this potentiality, we analyzed the differential expression of its two isoforms in human astrocytoma specimens, and the CD99 involved signaling pathways in glioma model U87MG cell line. CD99 was also analyzed in GBM molecular subtypes. Whole transcriptomes by RNA-Seq of CD99-siRNA, and functional in vitro assays in CD99-shRNA, that are found in U87MG cells, were performed. Astrocytoma of different malignant grades and U87MG cells only expressed CD99 isoform 1, which was higher in mesenchymal and classical than in proneural GBM subtypes. Genes related to actin dynamics, predominantly to focal adhesion, and lamellipodia/filopodia formation were down-regulated in the transcriptome analysis, when CD99 was silenced. A decrease in tumor cell migration/invasion, and dysfunction of focal adhesion, were observed in functional assays. In addition, a striking morphological change was detected in CD99-silenced U87MG cells, further corroborating CD99 involvement in actin cytoskeleton rearrangement. Inhibiting the overexpressed CD99 may improve resectability and decrease the recurrence rate of GBM by decreasing tumor cells migration and invasion.

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Multicomponent gold nano-glycoconjugate as a highly immunogenic and protective platform against Burkholderia mallei

npj Vaccines ◽

10.1038/s41541-020-00229-9 ◽

2020 ◽

Vol 5 (1) ◽

Author(s):

Daniel Tapia ◽

Javier I. Sanchez-Villamil ◽

Alfredo G. Torres

Keyword(s):

Rational Design ◽

Antibody Responses ◽

Effective Vaccine ◽

Antigen Delivery ◽

Burkholderia Mallei ◽

Protein Antigens ◽

Intrinsic Resistance ◽

Vaccine Platform ◽

A Genome

Abstract Burkholderia mallei (Bm) is a facultative intracellular pathogen and the etiological agent of glanders, a highly infectious zoonotic disease occurring in equines and humans. The intrinsic resistance to antibiotics, lack of specific therapy, high mortality, and history as a biothreat agent, prompt the need of a safe and effective vaccine. However, the limited knowledge of protective Bm-specific antigens has hampered the development of a vaccine. Further, the use of antigen-delivery systems that enhance antigen immunogenicity and elicit robust antigen-specific immune responses has been limited and could improve vaccines against Bm. Nanovaccines, in particular gold nanoparticles (AuNPs), have been investigated as a strategy to broaden the repertoire of vaccine-mediated immunity and as a tool to produce multivalent vaccines. To synthesize a nano-glycoconjugate vaccine, six predicted highly immunogenic antigens identified by a genome-wide bio- and immuno-informatic analysis were purified and coupled to AuNPs along with lipopolysaccharide (LPS) from B. thailandensis. Mice immunized intranasally with individual AuNP-protein-LPS conjugates, showed variable degrees of protection against intranasal Bm infection, while an optimized combination formulation (containing protein antigens OmpW, OpcP, and Hemagglutinin, along with LPS) showed complete protection against lethality in a mouse model of inhalational glanders. Animals immunized with different nano-glycoconjugates showed robust antigen-specific antibody responses. Moreover, serum from animals immunized with the optimized nano-glycoconjugate formulation showed sustained antibody responses with increased serum-mediated inhibition of adherence and opsonophagocytic activity in vitro. This study provides the basis for the rational design and construction of a multicomponent vaccine platform against Bm.

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Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens

PLoS Pathogens ◽

10.1371/journal.ppat.1009752 ◽

2021 ◽

Vol 17 (7) ◽

pp. e1009752

Author(s):

Jin-Tao Zou ◽

Hai-Ming Jing ◽

Yue Yuan ◽

Lang-Huan Lei ◽

Zhi-Fu Chen ◽

...

Keyword(s):

Protective Efficacy ◽

Antigen Delivery ◽

Protein Antigens ◽

Antigen Uptake ◽

Alpha Hemolysin ◽

Antibody Titers ◽

Candidate Antigen ◽

Draining Lymph Nodes

Highly immunogenic exotoxins are used as carrier proteins because they efficiently improve the immunogenicity of polysaccharides. However, their efficiency with protein antigens remains unclear. In the current study, the candidate antigen PA0833 from Pseudomonas aeruginosa was fused to the α-hemolysin mutant HlaH35A from Staphylococcus aureus to form a HlaH35A-PA0833 fusion protein (HPF). Immunization with HPF resulted in increased PA0833-specific antibody titers, higher protective efficacy, and decreased bacterial burden and pro-inflammatory cytokine secretion compared with PA0833 immunization alone. Using fluorescently labeled antigens to track antigen uptake and delivery, we found that HlaH35A fusion significantly improved antigen uptake in injected muscles and antigen delivery to draining lymph nodes. Both in vivo and in vitro studies demonstrated that the increased antigen uptake after immunization with HPF was mainly due to monocyte- and macrophage-dependent macropinocytosis, which was probably the result of HPF binding to ADAM10, the Hla host receptor. Furthermore, a transcriptome analysis showed that several immune signaling pathways were activated by HPF, shedding light on the mechanism whereby HlaH35A fusion improves immunogenicity. Finally, the improvement in immunogenicity by HlaH35A fusion was also confirmed with two other antigens, GlnH from Klebsiella pneumoniae and the model antigen OVA, indicating that HlaH35A could serve as a universal carrier protein to improve the immunogenicity of protein antigens.

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Antitumor Activity of Curcumin in Glioblastoma

International Journal of Molecular Sciences ◽

10.3390/ijms21249435 ◽

2020 ◽

Vol 21 (24) ◽

pp. 9435

Author(s):

Blake C. Walker ◽

Sandeep Mittal

Keyword(s):

Curcuma Longa ◽

Malignant Gliomas ◽

Standard Of Care ◽

Additional Treatment ◽

Treatment Modalities ◽

Molecular Signaling ◽

Current Standard ◽

Traditional Medicines ◽

Tumor Treating Fields

Current standard-of-care treatment for glioblastoma, the most common malignant primary central nervous system (CNS) tumor, consists of surgical resection followed by adjuvant chemotherapy and radiation (Stupp protocol), providing an overall median survival of 15 months. With additional treatment using tumor-treating fields (Optune® therapy, Novocure Ltd., Haifa, Israel), survival can be extended up to 20 months. In spite of significant progress in our understanding of the molecular pathogenesis, the prognosis for patients with malignant gliomas remains poor and additional treatment modalities are critically needed. Curcumin is a bright yellow pigment found in the rhizome of the widely utilized spice, turmeric (Curcuma longa). It has long been used in South Asian traditional medicines and has been demonstrated to have in vitro antioxidant, anti-inflammatory, and antiproliferative effects. Curcumin has been demonstrated to induce multiple cytotoxic effects in tumor cells including cell cycle arrest, apoptosis, autophagy, changes in gene expression, and disruption of molecular signaling. Additionally, curcumin has been shown to potentiate the effect of radiation on cancer cells, while exhibiting a protective effect on normal tissue. Curcumin’s positive safety profile and widespread availability make it a promising compound for future clinical trials for high-grade gliomas.

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Lysin CF-301 Demonstrates In Vitro Synergy with Conventional Antibiotics against Staphylococcus aureus

Open Forum Infectious Diseases ◽

10.1093/ofid/ofx163.908 ◽

2017 ◽

Vol 4 (suppl_1) ◽

pp. S370-S370 ◽

Cited By ~ 1

Author(s):

Karen Sauve ◽

Alena Jandourek ◽

Cara Cassino ◽

Raymond Schuch

Keyword(s):

Horse Serum ◽

Standard Of Care ◽

Therapeutic Benefit ◽

Synergistic Activity ◽

Current Standard ◽

Growth Interface ◽

Testing Medium ◽

Mrsa Strains ◽

Conventional Antibiotics

Abstract Background CF-301 is a novel, recombinantly-produced bacteriophage-derived lysin (cell wall hydrolase) and is the first agent of this class to enter clinical development in the US for the treatment of bacteremia including endocarditis due to S.aureus. This study evaluated the in vitro activity of CF-301 combined with each of 7 antistaphylococcal antibiotics including those considered to be current standard of care treatments for S.aureus bacteremia (daptomycin, vancomycin, oxacillin, nafcillin, and cefazolin) as well as linezolid and telavancin. Methods MICs for CF-301 were determined using a new AST medium for broth microdilution recently endorsed by the CLSI for use with CF-301. The testing medium consisted of cation-adjusted MHB supplemented with 25% horse serum and 0.5 mM DTT. Synergy was determined by checkerboard microdilution using the fractional inhibitory concentration index (FICI) for each combination in triplicate. For each antibiotic tested, an FIC mean was derived from each set of checkerboards by averaging 3 consecutive FIC values along the growth/no growth interface for each plate. Thus, 9 values were, used to generate the final mean. Synergy was defined as an FICI of ≤0.5; indifference was >0.5 to <2; and antagonism was ≥2. Each combination was examined against 10 MSSA and 10 MRSA strains. Results CF-301 synergized with daptomycin and vancomycin against each MSSA and MRSA strain, with FICI values between 0.254 and 0.5. Synergy was similarly observed against all 20 strains tested with oxacillin and nafcillin (FICI = 0.25–0.5); for the third β-lactam, cefazolin, synergy was observed with 17 strains (FICI = 0.75, for the remaining 3 strains). CF-301 synergized with televancin against 70% of the strains (FICI = 0.375–0.5), and was indifferent with the remainder (FICI = 0.625–1). CF-301 synergized with linezolid against 55% of the strains (FICI = 0.375–0.5), and was indifferent with the remainder (FICI = 0.625–0.75). Conclusion The broadly synergistic activity of CF-301 with conventional antistaphylococcal antibiotics against MSSA and MRSA suggests that CF-301 may afford therapeutic benefit by potentiating the activity antibiotics to treat serious infections for which there is an unmet medical need to improve outcomes. Disclosures K. Sauve, ContraFect Corp: Employee, Salary; A. Jandourek, ContraFect Corp: Employee, Salary; C. Cassino, ContraFect Corp: Employee, Salary; R. Schuch, ContraFect Corp: Employee, Salary

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An innovative cyanoacrylate device developed to improve the current standard of care for intravascular catheter securement

The Journal of Vascular Access ◽

10.1177/1129729819872881 ◽

2019 ◽

Vol 21 (3) ◽

pp. 293-299

Author(s):

Amanda Guido ◽

Sheng Zhang ◽

Cheng Yang ◽

Laura Pook

Keyword(s):

Cost Effective ◽

Standard Of Care ◽

Intravascular Catheter ◽

Current Standard ◽

Care Protocol ◽

Intravascular Catheters ◽

Intravascular Devices ◽

Strength And Durability

Introduction: Over one billion intravascular devices are used worldwide, annually. Due to the associated complications with these devices, the development of a reliable yet cost-effective securement technique is extremely important. The purpose of this study is to demonstrate the strength of a novel catheter securement cyanoacrylate for securing peripheral venous catheters, central venous catheters, peripherally inserted central catheters, and all other intravascular catheter types. Materials and methods: An unprecedented in vitro method was performed to quantify and compare the strength of a novel cyanoacrylate product when securing intravascular catheters inserted into prepared porcine skin. In vivo, canine subjects were used to implant various types of catheters. These catheters were secured with a novel catheter securement cyanoacrylate to test the strength and durability while undergoing simulated clinical stresses. Results: In vitro, the catheter securement cyanoacrylate demonstrated superior strength over conventional catheter securement methods as well as other known cyanoacrylates. The catheter securement cyanoacrylate demonstrated the ability to maintain superior strength for up to 7 days. In vivo, the catheter securement cyanoacrylate demonstrated the ability to withstand five weight tugs per hour for a 3-h duration, alone, while securing three types of catheters in canine subjects. Conclusion: This is one of the first studies to provide quantitative data to support the use of cyanoacrylate for intravascular catheter securement. The results from this research suggest that the novel catheter securement cyanoacrylate can be a simple and cost-effective catheter securement device that can improve the current health care protocol for intravascular catheterization.

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Radiological response patterns in the phase 2 STELLAR trial of TTFields with chemotherapy for first-line treatment of malignant pleural mesothelioma (MPM).

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.8551 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. 8551-8551

Author(s):

Federica Grosso ◽

Giovanni Luca Ceresoli

Keyword(s):

Disease Progression ◽

Electric Fields ◽

Combined Therapy ◽

Response Duration ◽

Standard Of Care ◽

First Line ◽

Current Standard ◽

Median Response ◽

Tumor Treating Fields

8551 Background: Tumor Treating Fields (TTFields) are an anti-mitotic, regional treatment modality, utilizing low intensity alternating electric fields delivered non-invasively to the tumor using a portable, medical device. TTFields have significantly extended survival of patients with glioblastoma. In-vitro, human MPM cells were highly susceptible to TTFields. In the STELLAR study, patients with unresectable MPM treated with first line chemotherapy in combination with TTFields had a significantly higher median overall survival compared to historical controls (18.2 vs. 12.1 months). We report on analysis of radiological data from STELLAR patients whose tumors responded while receiving the combined therapy. Methods: The STELLAR trial accrued 80 patients with unresectable, previously untreated mesothelioma. Patients were treated with continuous 150 kHz TTFields (>18h/day) in combination with pemetrexed and cisplatin or carboplatin (at standard dosing). Inclusion criteria included ECOG PS of 0-1, pathologically proven mesothelioma and at least one measurable lesion according to modified RECIST criteria. Patients were followed q3w (CT scan q6w) until disease progression. Radiological assessments were done at each study site. Results: Partial responses (PRs) were seen in 40.3% of evaluable patients and clinical benefit (PR+SD) was seen in 97.2% of these patients. The median time between treatment start and PR was 1.8 months (range: 1.4-4.4 months). All patients presenting with PR during the STELLAR study had continuous reduction in the total sum of lesion diameters, suggesting no initial / pseudo-progression. 83% of the patients who responded to the combined therapy finally had disease progression within a median response duration of 5.7 months (range: 1.4-13 months), per Kaplan-Meier Estimator. One patient did not progress for more than 27 months. Conclusions: The STELLAR study met its primary endpoint of significant survival extension for previously untreated mesothelioma patients. Response rates were similar to the ones reported for the current standard of care treatment, but lasted longer with the addition of TTFields. Clinical trial information: NCT02397928.

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The PI3K Inhibitor GDC-0941 Synergizes with Standard of Care Therapies to Induce Growth Inhibition and Apoptosis of Multiple Myeloma Cells.

Blood ◽

10.1182/blood.v114.22.3788.3788 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3788-3788

Author(s):

Veerendra Munugalavadla ◽

Leanne Berry ◽

Changchun Du ◽

Sanjeev Mariathasan ◽

Dion Slaga ◽

...

Keyword(s):

Multiple Myeloma ◽

Research Funding ◽

Single Agent ◽

Standard Of Care ◽

Rational Approach ◽

Current Standard ◽

Employment Equity ◽

Specific Flow ◽

Equity Ownership

Abstract Abstract 3788 Poster Board III-724 Multiple myeloma (MM) is a malignancy characterized by clonal expansion and accumulation of long-lived plasma cells within the bone marrow. Phosphatidylinositol 3' kinase (PI3K) -mediated signaling is frequently dysregulated in cancer and controls fundamental cellular functions such as cell migration, growth, survival and development of drug resistance in many cancers, including MM, and therefore represents an attractive therapeutic target. Here, we demonstrate in vitro, that a potent and selective pan-isoform PI3Kinhibitor, GDC-0941, modulates the expected pharmacodynamic markers, inhibits cell-cycle progression and induces apoptosis; overcomes resistance to apoptosis in MM cells conferred by IGF-1 and IL-6; and is additive or synergistic with current standard of care drugs including dexamethasone, melphalan, lenolidamide and bortezomib. In cell lines we find sensitivity to GDC-0941 is positively correlated with pathway activation as determined by phospho-AKT-specific flow-cytometry and Western-blot analysis. Preliminary results indicate apoptosis of MM cells is correlated with increased expression of the proapoptotic BH3-only protein BIM; mechanisms of increased apoptosis in MM will be further explored and an update presented. We further extend these in vitro findings to show that GDC-0941 has activity as a single agent in vivo and combines well with standard of care agents in several murine xenograft models to delay tumor progression and prolong survival. Our results suggest that GDC-0941 may combine well with existing therapies, providing a framework for the clinical use of this agent, and a rational approach to improving the efficacy of myeloma treatment. Disclosures: Munugalavadla: Genentech: Employment, Patents & Royalties. Berry:Genentech: Employment, Patents & Royalties. Du:Genentech, Inc.: Employment, Equity Ownership. Mariathasan:Genentech: Employment, Patents & Royalties. Slaga:Genentech: Employment, Patents & Royalties. Sun:Genentech Inc.: Employment. Chesi:Genentech, Inc.: Consultancy; Amgen: Consultancy; Celgene: Consultancy; Merck: Research Funding. Bergsagel:Genentech: Consultancy; Amgen: Consultancy; Celgene: Consultancy; Merck: Research Funding. Ebens:Genentech, Inc.: Employment, Equity Ownership, Patents & Royalties.

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