Michael Andrew Luzuriaga

The increasing rate of resistance of bacterial infection against antibiotics requires next generation approaches to fight potential pandemic spread. The development of vaccines against pathogenic bacteria has been difficult owing, in part, to the genetic diversity of bacteria. Hence, there are many potential target antigens and little a priori knowledge of which antigen/s will elicit protective immunity. The painstaking process of selecting appropriate antigens could be avoided with whole-cell bacteria; however, whole-cell formulations typically fail to produce long-term and durable immune responses. These complications are one reason why no vaccine against any type of pathogenic E. coli has been successfully clinically translated. As a proof of principle, we demonstrate a method to enhance the immunogenicity of a model pathogenic E. coli strain by forming a slow releasing depot. The E. coli strain CFT073 was biomimetically mineralized within a metal–organic framework (MOF). This process encapsulates the bacteria within 30 min in water and at ambient temperatures. Vaccination with this formulation substantially enhances antibody production and results in significantly enhanced survival in a mouse model of bacteremia compared to standard inactivated formulations.

Vaccines are considered one of the most significant medical advancements in human history, as they have prevented hundreds of millions of deaths since their discovery; however, modern travel permits disease spread at unprecedented rates, and vaccine shortcomings like thermal sensitivity and required booster shots have been made evident by the COVID‐19 pandemic. Approaches to overcoming these issues appear promising via the integration of vaccine technology with biomaterials, which offer sustained‐release properties and preserve proteins, prevent conformational changes, and enable storage at room temperature. Sustained release and thermal stabilization of therapeutic biomacromolecules is an emerging area that integrates material science, chemistry, immunology, nanotechnology, and pathology to investigate different biocompatible materials. Biomaterials, including natural sugar polymers, synthetic polyesters produced from biologically derived monomers, hydrogel blends, protein–polymer blends, and metal–organic frameworks, have emerged as early players in the field. This overview will focus on significant advances of sustained release biomaterial in the context of vaccines against infectious disease and the progress made towards thermally stable “single‐shot” formulations.This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Infectious Disease

Artificial native-like lipid bilayer systems constructed from phospholipids assembling into unilamellar liposomes allow the reconstitution of detergent-solubilized transmembrane proteins into supramolecular lipid-protein assemblies called proteoliposomes, which mimic cellular membranes. Stabilization of these complexes remains challenging because of their chemical composition, the hydrophobicity and structural instability of membrane proteins, and the lability of interactions between protein, detergent, and lipids within micelles and lipid bilayers. In this work we demonstrate that metastable lipid, protein-detergent, and protein-lipid supramolecular complexes can be successfully generated and immobilized within zeolitic-imidazole framework (ZIF) to enhance their stability against chemical and physical stressors. Upon immobilization in ZIF bio-composites, blank liposomes, and model transmembrane metal transporters in detergent micelles or embedded in proteoliposomes resist elevated temperatures, exposure to chemical denaturants, aging, and mechanical stresses. Extensive morphological and functional characterization of the assemblies upon exfoliation reveal that all these complexes encapsulated within the framework maintain their native morphology, structure, and activity, which is otherwise lost rapidly without immobilization.

Solid-state thermoelastic behavior—a sudden exertion of an expansive or contractive physical force following a temperature change and phase transition in a solid-state compound—is rare in organic crystals, few are reversible systems, and most of these are limited to a dozen or so cycles before the crystal degrades or they reverse slowly over the course of many minutes or even hours. Comparable to thermosalience, wherein crystal phase changes induce energetic jumping, thermomorphism produces physical work via consistent and near-instantaneous predictable directional force. In this work, we show a fully reversible thermomorphic actuator that is stable at room temperature for multiple years and is capable of actuation for more than 200 cycles at near-ambient temperature. Specifically, the crystals shrink to 90% of their original length instantaneously upon heating beyond 45 °C and expand back to their original length upon cooling below 35 °C. Furthermore, the phase transition occurs instantaneously, with little obvious hysteresis, allowing us to create real-time actuating thermal fuses that cycle between on and off rapidly.

Solid state thermosalience—a sudden exertion of an expansive or contractive physical force following a temperature change in a solid state compound—is rare, few are reversible systems, and most of these are limited to a dozen or so cycles before the crystal degrades or they reverse slowly over the course of many minutes or even hours. In this work, we show a fully reversible actuator that is stable at room temperature for multiple years and is capable of actuation for more than two hundred cycles at near ambient temperature. Specifically, the crystals shrink to 90% of its original length instantaneously upon heating beyond 45 °C and expands back to its original length upon cooling below 35 °C. This temperature regime is important because it occurs around physiologically important temperatures. Furthermore, the phase transition occurs instantaneously, with little obvious hysteresis, allowing us to create real-time actuating thermal fuses that cycle between on and off rapidly.

Virus like particles (VLPs) are non-infectious multifunctional nanocarriers with the pathogenic-like architecture of viruses. They can serve as a safe platform for precise functionalization and immunization, which provides benefits in a wide range of biomedical applications. In this work, we describe the development of novel immunophotothermal agent for combinatorial photothermal ablation and immunotherapy based on VLP of bacteriophage Qβ. The design was based on covalent conjugation of 212 water soluble near-infrared absorbing croconium dyes to lysine residues located on the surface of Qβ, which turned it to a powerful NIR-absorber with photothermal efficiencies exceeding that of gold nanostructures. This PhotoPhage system generates heat upon 808 nm NIR laser radiation and causes significant cellular cytotoxicity that prevents the progression of primary tumors in mice. It is found that PhotoPhage not only acts as a PTT agent that initiates anti-tumor immune response, but also simultaneously acts as an immunoadjuvant that promotes maturation of dendritic cells, triggers T lymphocyte cells (CD4+, CD8+) and reduces suppressive T regulatory cells leading to effective suppression of primary tumors, reducing lung metastases, and increasing survival time.

Artificial native-like lipid bilayer systems constructed from phospholipids assembling into unilamellar liposomes allow the reconstitution of detergent-solubilized transmembrane proteins into supramolecular lipid-protein assemblies called proteoliposomes, which mimic cellular membranes. Stabilization of these complexes remains challenging because of their chemical composition, the hydrophobicity and structural instability of membrane proteins, and the lability of interactions between protein, detergent, and lipids within micelles and lipid bilayers. In this work we demonstrate that metastable lipid, protein-detergent, and protein-lipid supramolecular complexes can be successfully generated and immobilized within zeolitic-imidazole framework (ZIF) to enhance their stability against chemical and physical stressors. Upon immobilization in ZIF bio-composites, blank liposomes, and model transmembrane metal transporters in detergent micelles or embedded in proteoliposomes resist elevated temperatures, exposure to chemical denaturants, aging, and mechanical stresses. Extensive morphological and functional charac- terization of the assemblies upon exfoliation reveal that all these complexes encapsulated within the framework maintain their native morphology, structure, and activity, which is otherwise lost rapidly without immobilization.

Urinary tract infection (UTI) is the most common adult bacterial infections worldwide, with ~80% of all uncomplicated UTI caused by uropathogenic E. coli (UPEC). While anyone can develop a UTI, women in their 20’s and postmenopausal women are at particularly high risk—indeed, half of all women will develop a UTI in their lifetime. Even with antibiotic therapy, 25-50% of patients will experience recurrence within a year.¹ Initial and subsequent UTIs can alter bladder epithelial cell barrier function, which in turn creates an environment more favorable to subsequent infection.² This results in recurrent UTI, which requires long-term antimicrobial therapy and can reoccur in a patient indefinitely, causing a lifetime of pain, discomfort, and an elevated risk of septicemic death.³ Unfortunately, antibiotic resistance makes treating UTIs less effective. Indeed, 10–25% of uncomplicated UTI patient isolates are resistant to trimethoprim/sulfamethoxazole (TMP/SMX)—the standard-of-care antibiotic regime for UTI.^{4 , 5} Alarmingly, >70% of postmenopausal recurrent UTI patients are either allergic or resistant to TMP-SMX.⁵ Given the ubiquity of these infections and the high likelihood of recurrence, the societal cost is estimated to be over $5 billion each year in the US alone.⁶ A prophylactic or therapeutic vaccine would be an ideal first line of defense against UTIs for high risk groups, but no FDA-approved vaccines against UTI—or indeed any pathogenic E. coli infection—exist.⁷

Solid state thermosalience—a sudden exertion of an expansive or contractive physical force following a temperature change in a solid state compound—is rare, few are reversible systems, and most of these are limited to a dozen or so cycles before the crystal degrades or they reverse slowly over the course of many minutes or even hours. In this work, we show a fully reversible actuator that is stable at room temperature for multiple years and is capable of actuation for more than two hundred cycles at near ambient temperature. Specifically, the crystals shrink to 90% of its original length instantaneously upon heating beyond 45 °C and expands back to its original length upon cooling below 35 °C. This temperature regime is important because it occurs around physiologically important temperatures. Furthermore, the phase transition occurs instantaneously, with little obvious hysteresis, allowing us to create real-time actuating thermal fuses that cycle between on and off rapidly.

Artificial native-like lipid bilayer systems constructed from phospholipids assembling into unilamel- lar liposomes allow the reconstitution of detergent-solubilized transmembrane proteins into supramolecular lipid-protein assemblies called proteoliposomes, which mimic cellular membranes. Stabilization of these com- plexes remains challenging because of their chemical composition, the hydrophobicity and structural instabil- ity of membrane proteins, and the lability of interactions between protein, detergent, and lipids within micelles and lipid bilayers. In this work we demonstrate that metastable lipid, protein-detergent, and protein-lipid su- pramolecular complexes can be successfully generated and immobilized within zeolitic-imidazole framework- 8 (ZIF-8) to enhance their stability against chemical and physical stressors. Upon immobilization in ZIF-8 bio- composites, blank liposomes, and model transmembrane metal transporters in detergent micelles or embed- ded in proteoliposomes resist elevated temperatures, exposure to chemical denaturants, aging, and mechanical stresses. Extensive morphological and functional characterization of the assemblies upon exfoliation reveal that all these complexes encapsulated within the framework maintain their native morphology, structure, and activity, which is otherwise lost rapidly without immobilization.