Thomas J Anchordoquy

Thomas J Anchordoquy
University of Colorado | UCD · Department of Pharmaceutical Sciences

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107
Publications
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Introduction

Publications

Publications (107)
Article
It is becoming increasingly clear that the intravenous administration of nanoparticles elicits an immune response that compromises delivery efficiency and can be life threatening. This study investigated both the systemic and tissue-level cytokine response to repeat administration of lipoplexes coated with either lactose or PEG. We report that bloo...
Article
Studies have suggested imatinib mesylate (ImM) as a potential treatment for systemic lupus erythematosus nephritis (SLEN). However, ImM has limited renal excretion. The goal of the current research was to develop an ImM containing nanoformulation, conduct studies to evaluate pharmacokinetics, and determine whether kidney deposition can be enhanced...
Article
Full-text available
As mRNA vaccines became the frontrunners in late-stage clinical trials to fight the COVID-19 pandemic, challenges surrounding their formulation and stability became readily apparent. In this commentary, we first describe company proposals, based on available public information, for the (frozen) storage of mRNA vaccine drug products across the vacci...
Article
Nanoparticle-mediated drug delivery has long utilized PEGylation as a mechanism for reducing uptake by the reticuloendothelial system and extending circulation lifetimes. However, studies over the past two decades have established that immune responses to PEG can promote clearance upon repeat injection and elicit life-threatening anaphylactic react...
Article
Aim: Conventional conjugation reactions often involve the use of activated PEG as a linker, but concerns about PEG-mediated reduction in intracellular delivery and enhanced immunogenicity have generated interest in developing methods that eliminate the need for a PEG linker. Materials & methods: Reaction conditions were identified that specifica...
Article
Full-text available
Many pharmaceuticals must be administered intravenously due to their poor oral bioavailability. In addition to issues associated with sterility and inconvenience, the cost of repeated infusion over a 6-week course of therapy costs the health care system tens of billions of dollars per year. Attempts to improve oral bioavailability have traditionall...
Article
Decades of research into improving drug delivery to tumors has documented uptake of particulate delivery systems by resident macrophages in the lung, liver, and spleen, and correlated short circulation times with reduced tumor accumulation. An implicit assumption in these studies is that nanoparticles present in the blood are available for distribu...
Article
Glioblastoma multiforme (GBM) is a devastating tumour with abysmal prognoses. We desperately need novel approaches to understand GBM biology and therapeutic vulnerabilities. Extracellular vesicles (EVs) are membrane-enclosed nanospheres released locally and systemically by all cells, including tumours, with tremendous potential for intercellular co...
Chapter
The ability of cholesterol to impart stability and serum-resistance to lipid nanoparticles has been well-known for several decades. Thus, the use of cholesterol offers an alternative to utilizing the increasingly problematic “stealth” technology. More recently, the ability of high amounts of cholesterol to form nanodomains within lipid nanoparticle...
Article
Remarkable progress has recently been made in the synthesis and characterization of engineered nanoparticles for imaging and treatment of cancers, resulting in several promising candidates in clinical trials. Despite these advances, clinical applications of nanoparticle-based therapeutic/imaging agents remain limited by biological, immunological, a...
Article
Full-text available
Background: Agents targeting HSP90 and GRP94 are seldom tested in stressed contexts such as heat shock (HS) or the unfolded protein response (UPR). Tumor stress often activates HSPs and the UPR as pro-survival mechanisms. This begs the question of stress effects on chemotherapeutic efficacy, particularly with drugs targeting chaperones such as HSP9...
Article
Repetitive administration is routinely used to maintain therapeutic drug levels, but previous studies have documented an accelerated blood clearance of some lipid-based delivery systems under these conditions. To assess the effect of repetitive administration, non-PEGylated lipoplexes (+/- = 0.5) were administered four times via tail vein injection...
Article
The relevance of using nude mouse models for evaluating drug delivery to human tumors has recently been questioned by numerous researchers. While the immune response is known to play a critical role in cancer, this study assesses the effect of using immunocompromised “nude” mice on drug delivery. By inoculating both nude and immunocompetent mice wi...
Article
Full-text available
Brain tumors such as high grade gliomas are among the deadliest forms of human cancers. The tumor environment is subject to a number of cellular stressors such as hypoxia and glucose deprivation. The persistence of the stressors activates the unfolded proteins response (UPR) and results in global alterations in transcriptional and translational act...
Article
Background: The vast majority of studies investigating gene delivery have utilized cationic delivery vehicles, but anionic nanoparticles can also possess high transfection activity, and offer significant benefits in terms of ease of preparation and reduced toxicity. Results: Our study on lipoplexes possessing cholesterol nanodomains demonstrates...
Article
Full-text available
Background: Glioma-related immunosuppression is well documented; however, the mechanisms of suppression are not fully understood. Here we explore a role for glioma extracellular vesicles (EVs) as a means of immune modulation. Methods: Healthy donor peripheral blood mononuclear cells (PBMCs) were incubated with mitogenic stimuli and various conce...
Chapter
The ability to deliver nucleic acids in vivo offers the promise to develop potent pharmaceutical medicines (e.g., vaccines) to treat many human diseases such as cancer, AIDS, and others that are currently considered incurable. It is well recognized that nonviral gene delivery systems exhibit high instability as liquid formulations. Therefore, the d...
Article
In cancer patients, granulocytic myeloid derived suppressor cells (G-MDSCs) expand in number, infiltrating tumor and lymphatic tissues where they suppress an anti-tumor immune response. We report here the development of a liposomal drug delivery system that selectively targets G-MDSCs. The liposomes form a disulfide bond with activated complement C...
Article
Unlabelled: The protein corona that forms around nanoparticles in vivo is a critical factor that affects their physiological response. The potential to manipulate nanoparticle characteristics such that either proteins advantageous for delivery are recruited and/or detrimental proteins are avoided offers exciting possibilities for improving drug de...
Article
Full-text available
The use of exosomes as a drug delivery vehicle has gained considerable interest. To establish if exosomes could be utilized effectively for drug delivery, a better understanding of their in vivo fate must be established. Through comparisons to liposomal formulations, which have been studied extensively for the last thirty years, we were able to mak...
Article
Because aqueous liposomal formulations containing multiply unsaturated lipids are susceptible to chemical degradation, these formulations are often lyophilized. Despite their limited chemical stability, interest in the use of multiply unsaturated lipids to promote intracellular delivery has increased considerably in recent years. The goal of the cu...
Article
Cationic reagents are commonly used to facilitate DNA delivery, and transfection experiments are typically initiated in cell culture where the optimal charge ratio is determined. While transfection rates are often enhanced at higher +/- charge ratios, the cellular toxicity associated with the greater amounts of cationic components at elevated charg...
Article
Full-text available
A method for conjugation of ligands to the surface of exosomes was developed using click chemistry. Copper-catalyzed azide alkyne cycloaddition (click chemistry) is ideal for biocojugation of small molecules and macromolecules to the surface of exosomes, due to fast reaction times, high specificity, and compatibility in aqueous buffers. Exosomes cr...
Article
Conjugation of polyethylene glycol (PEG) to therapeutics has proven to be an effective approach to increase the serum half-life. However, the increased use of PEGylated therapeutics has resulted in unexpected immune-mediated side-effects. There are claims that these are caused by anti-PEG antibodies inducing rapid clearance. These claims are howeve...
Article
Small endogenous vesicles called exosomes are beginning to be explored as drug delivery vehicles. The in vivo targets of exosomes are poorly understood; however, they are believed to be important in cell-to-cell communication and may play a prominent role in cancer metastasis. We aimed to elucidate whether cancer derived exosomes can be used as dru...
Article
Full-text available
Unlabelled: Current liposomal gene delivery systems predominately utilize cationic lipids, which efficiently bind and deliver DNA plasmid, but also result in nonspecific gene expression in lung and liver tissue. To improve specificity, a two-component delivery strategy employing neutral liposomes was used to target breast cancers positive for the...
Article
The objective of our research is to develop a liposomal gene delivery system that specifically targets potent therapeutic genes to breast cancers which are positive for the human epidermal growth receptor 2 (Her-2). Current liposomal gene delivery systems predominately utilize cationic lipids, which efficiently bind and deliver plasmid DNA, but als...
Article
Full-text available
Polyethylene glycol (PEG) is widely utilized in drug delivery and nanotechnology due to its reported “stealth” properties and biocompatibility. It is generally thought that PEGylation allows particulate delivery systems and biomaterials to evade the immune system and thereby prolong circulation lifetimes. However, numerous studies over the past dec...
Article
Full-text available
The objective of a systemically administered cancer gene therapy is to achieve gene expression that is isolated to the tumor tissue. Unfortunately, viral systems have strong affinity for the liver, and delivery from non-viral cationic systems often results in high expression in the lungs. Non-specific delivery to these organs must be overcome if tu...
Article
The use of lipoplexes for the intracellular delivery of nucleic acids typically involves the optimization of several parameters that are known to affect delivery. Researchers commonly vary charge ratio, and often incorporate different amounts of helper lipids (e.g., cholesterol) to optimize formulations for transfection in cell culture and in vivo....
Conference Paper
Full-text available
Brain diseases such as cancers, neurodegenerative disorders, or trauma are frequently diagnosed with imaging modalities and sometimes with intracranial biopsies. Treatment response is similarly monitored, along with clinical indications. While these technologies provide important windows into the disease state, they fail to provide us a detailed mo...
Article
As liquid liposomal formulations are prone to chemical degradation and aggregation, these formulations often require freeze drying (e.g. lyophilization) to achieve sufficient shelf-life. However, liposomal formulations may undergo oxidation during lyophilization and/or during prolonged storage. The goal of the current study was to characterize the...
Article
Background: The formation of cholesterol domains in lipoplexes has been associated with enhanced serum stability and transfection rates both in cell culture and in vivo. Results: This study utilizes the ability of saturated phosphatidylcholines to promote the formation of cholesterol domains at much lower cholesterol contents than have been util...
Article
Full-text available
Medulloblastomas are the most prevalent malignant pediatric brain tumors. Survival for these patients has remained largely the same for approximately 20 years, and our therapies for these cancers cause significant health, cognitive, behavioral and developmental sequelae for those who survive the tumor and their treatments. We obviously need a bette...
Data
Extended list of Top Network Functions/Biofunctions: “Diseases and Disorders” from IPA Core Analysis. This list encompasses the top 72 categories with scores above the threshold for significance. “Threshold” indicates the minimum significance level (scored as –log [p value] from Fisher’s exact test, set here at 1.25). (TIF)
Data
Results of D283MED exosome proteomics. Table lists proteins identified by mass spectrometry or Western blot analyses. Gene/protein IDs, symbols, and names are presented, along with peptide count, source of identification (MS or Western blot), proteins’ predicted subcellular localizations and putative functions, and presence in the ExoCarta database...
Data
Extended list of the Top Network Functions “Top Canonical Pathways” from IPA Core Analysis. This list encompasses the top 28 categories with scores above the threshold for significance. “Threshold” indicates the minimum significance level (scored as –log [p value] from Fisher’s exact test, set here at 1.25). “Ratio” indicates the number of molecule...
Data
Top Network Functions “Top Toxicology Functions and Lists” from IPA Core Analysis. (A) shows the top 12 significantly-scoring toxicology functions derived from IPA analyses; (B) shows the top 5 toxicology lists. The statistically significant threshold is defined as in Figures S1 and S2. (TIF)
Article
Bleomycin is a membrane impermeable chemotherapeutic agent that is relatively innocuous extracellularly but highly cytotoxic when delivered directly to the cytoplasm. We report on the development of a liposome delivery system that targets Her-2 overexpressing breast cancer cells and breaches the endosomal barrier, delivering bleomycin to the cytopl...
Article
Targeted gene delivery provides enormous potential for clinical treatment of many incurable diseases. Liposomes formulated with targeting ligands have been tested extensively both in vitro and in vivo, and many studies have strived to identify more efficacious ligands. However, the environment of the ligand within the delivery vehicle is generally...
Article
The development of plasmid DNA as a pharmaceutical requires that integrity (i.e., supercoil content) be monitored as part of quality control. The standard method of determining supercoil content is gel electrophoresis followed by staining and imaging, which is complicated by a variety of factors. Previously described chromatographic methods used to...
Article
Particle-mediated epidermal delivery (PMED) of small quantities of DNA (0.5-4.0 μg) has been reported to both induce an immune response and protect against disease in human subjects. In order for the PMED of DNA to be a viable technique for vaccination, the adsorbed DNA must be stable during shipping and storage. Here, we report that the storage st...
Article
Strict US Food and Drug Administration regulations on contamination levels for DNA therapeutics acceptable for human use complicate the manufacturing process. This study aims to improve therapeutic production through the investigation of the molecular effects of hydrodynamic forces encountered during processing. Results suggest that the strain rate...
Article
Full-text available
PEGylated components have been widely used to reduce particle aggregation in serum and extend circulation lifetime for lipid- and polymer-based gene-delivery systems. However, PEGylation is known to interfere with cell interaction and intracellular trafficking, resulting in decreased biological activity. In the present study, the effect of choleste...
Article
Aerosolization techniques for delivery of gene therapies to the lungs decrease activity of these treatments. The low transfection is attributed to loss in molecular integrity. Electrosprays can aerosolize DNA without structural loss. Electrospray affects on human pulmonary cells are unknown. This study is to assess toxicity, inflammatory response,...
Article
The stability of pharmaceuticals and biological products is a major challenge for the industry. While some preparations have adequate stability as a liquid formulation, many biological molecules must be dried in order to achieve shelf-lives compatible with a commercial product. In addition, the technology for conducting freeze-drying and sample ana...
Article
The ability to deliver nucleic acids (e.g., plasmid DNA, antisense oligonucleotides, siRNA) offers the potential to develop potent vaccines and novel therapeutics. However, nucleic acid-based therapeutics are still in their early stages as a new category of biologics. The efficacy of nucleic acids requires that these molecules be delivered to the i...
Article
Targeted gene delivery offers immense potential for clinical applications. Liposomes decorated with targeting ligands have been extensively used for both in vitro and in vivo gene delivery. Lipoplexes with high cholesterol content that result in cholesterol domain formation within the complexes have been shown to exhibit enhanced transfection in vi...
Article
The stability of nonviral vectors during freeze-drying has been well-studied, and it has been established that sugars can protect lipoplexes during freeze-drying. However low levels of damage are often observed after freeze-drying, and this damage is more evident in dilute lipoplex preparations. By investigating the stability of lipoplexes after ea...
Article
The purpose of this study is to investigate the effects of moisture content on the storage stability of freeze-dried lipoplex formulations. DC-Cholesterol: DOPE (dioleoyl phosphatidylethanolamine)/plasmid DNA lipoplexes were prepared at a 3-to-2 DC-Cholesterol(+) to DNA(-) molar ratio and lyophilized prior to storing at room temperature, 40, and 60...
Article
Supercoiled DNA plasmids were exposed in the frozen state to high-energy electrons. Surviving supercoiled molecules were separated from their degradation products (e.g., open circle and linear forms) by agarose gel electrophoresis and subsequently quantified by staining and image analysis. Complex survival curves were analyzed using radiation targe...
Article
It has been shown that degradation of lipid/DNA complexes (lipoplexes) continues in the dried state during storage. The goal of this study was to evaluate the ability of various strategies to minimize the formation of reactive oxygen species (ROS) in lyophilized lipoplexes during storage, including metal removal from reagents, air displacement, and...
Article
The freeze-drying properties of gelatin nanoparticles were investigated with the goal of providing practicable nanoparticle formulations for in vitro applications or clinical studies. Various excipients and rehydration protocols were assessed, and gelatin nanoparticles loaded with oligonucleotides were successfully freeze-dried and rehydrated. An N...
Article
Isoprene is emitted from the leaves of some plants. It was recently reported that exogenous isoprene delays the onset of leaf damage during controlled increases in leaf temperature (Singsaas et al. Plant Physiology 115: 1413–1420 [1997[17). Thylakoid membranes are presumed to be the site of action based upon isoprene's hydrophobicity, production in...
Article
The interaction between the cationic lipid DOTAP and cholesterol is examined in high cholesterol formulations by differential scanning calorimetry (DSC). Preparation of liposomes above 66 mol% cholesterol results in formulations that exhibit a calorimetric transition for anhydrous cholesterol at 38-40 degrees C. The enthalpy of this transition prog...
Article
The presence of trace amounts of metal ions in nonviral vector formulations can significantly affect the stability of lipid/DNA complexes (lipoplexes) during acute freeze-drying. The goal of the present study was to evaluate the generation of reactive oxygen species (ROS) in dried formulations of lipoplexes and in their individual components (lipid...
Article
While clozapine is the acknowledged superior pharmacotherapeutic for the treatment of schizophrenia, the side effect profile, which includes potentially fatal complications, limits its usefulness. Central administration of clozapine directly into the brain could circumvent many of the side effect issues due to the dramatic reduction in dose and the...
Article
To evaluate the in vivo biodistribution and expression of DOTAP-Chol/DNA complexes (lipoplexes) with different in vitro serum stability, quantitative real-time PCR, in vitro luciferase expression and whole body luminescence imaging were used. In general, less tissue biodistribution, lower luciferase expression and whole body luminescence were obser...
Article
Frozen solutions of high molecular weight circular DNA were irradiated with high-energy electrons. Molecules that survived the radiation exposure were detected by structural and functional techniques and were analyzed using radiation target theory. Radiation damage (as a loss of function) was not observed in a distant portion in the same chain, ind...
Article
The preservation of DNA is of wide interest to scientists in disparate fields from biorepository management to pharmaceutical sciences. Although the term "preservation" is used by all these fields and refers to the maintenance of chemical and physical integrity of the DNA molecule, it should not be surprising that the perspectives of scientists fro...
Article
Oxidation reactions represent an important degradation pathway of nucleic acid-based pharmaceuticals. To evaluate the role of metal contamination and chelating agents in the formation of reactive oxygen species (ROS) during lyophilization, ROS generation and the stability of lipid/DNA complexes were investigated. Trehalose-containing formulations w...
Chapter
Introduction to BioprocessingReview of Molecular StructureManufacturing ConcernsExample Process—AtomizationSeparation TechniquesChapter SummaryReferences
Article
The regulatory roles fulfilled by polyamines by governance of chromatin structure are made possible by their strong association with cellular DNA, and hence by their ability to modulate DNA structure and function. Towards this end, it is crucial to understand the manifestation of sequence-dependent polyamine binding at the secondary and tertiary st...
Article
Several new technologies based upon ultrasound technology have been proposed as a method to enhance the delivery of genetic therapeutics. Using ultrasonic nebulization and a well-established method to quantitatively monitor transient cavitation, this study investigates the extent and factors influencing the degradation of DNA. Results from our stud...
Article
Genetic therapeutics show great promise toward the treatment of illnesses associated with the lungs; however, current methods of delivery such as jet and ultrasonic nebulization decrease the activity and effectiveness of these treatments. Extremely low transfection rates exhibited by non-complexed plasmid DNA in these nebulizers have been primarily...
Article
Jet nebulization is a common technique by which gene therapies can be delivered to the lungs, however current research has shown a reduction in activity and effectiveness of these treatments when not complexed with cationic agents. To date, a systematic investigation of chemical and mechanical degradation pathways has yet to be undertaken to identi...
Article
Stabilization of nonviral vectors during freezing and drying requires formulation with protective excipients such that transfection rates and physical characteristics are maintained upon reconstitution. While many studies have demonstrated the ability of disaccharides (e.g., sucrose) to effectively protect nonviral vectors during lyophilization, th...
Article
The importance of understanding the dynamics of DNA condensation is inherent in the biological significance of DNA packaging in cell nuclei, as well as for gene therapy applications. Specifically, the role of ligand hydrophobicity in DNA condensation has received little attention. Considering that only multivalent cations can induce true DNA conden...
Article
The development of nonviral vectors as commercial therapeutics will require formulations that are sufficiently stable to allow shipping and storage for prolonged periods. Given the well-known instability of these systems as aqueous suspensions, it would be desirable to develop lyophilized formulations that are resistant to shipping stress and can b...
Article
Plasmid DNA encoding the green lantern protein was ion-paired with 1,2-dioleoyl, 3-trimethylammonium propane (DOTAP) at a (+/-) charge ratio of (1:1) to form a hydrophobic ion-pair (HIP) complex using the Bligh and Dyer method, and transferred into methylene chloride. Precipitation with a compressed antisolvent (PCA) was then employed to encapsulat...
Article
It is well known that excipients are required to protect nonviral vectors during the lyophilization process. The goal of this study is to describe the stability of lyophilized nonviral vector preparations on pharmaceutically relevant timescales and provide insight into the factors that govern long-term stability of vectors in the dried state. Lipid...
Article
To evaluate the role of lipid charge density in the serum stability of DOTAP-Chol/DNA complexes (lipoplexes), lipid-DNA interactions, extent of aggregation, supercoil content, and in vitro transfection efficiency of lipoplexes were investigated. In general, higher serum concentration destabilized, and increasing molar charge ratio of DOTAP to negat...