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Living Microrobot for Active Therapeutic Delivery to Treat Severe Pulmonary Infections

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NIAID - National Institute of Allergy and Infectious Diseases

Project Summary Multidrug-resistant (MDR) lower respiratory tract infections represent the single leading cause of infectious disease-associated mortality in the United States. Particularly worrisome trends are being observed in the case of ventilator-associated pneumonia (VAP), which affects vulnerable patient populations in intensive care units (ICUs). Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus (MRSA) are the most common causative agents in global epidemiology of VAP, and they are becoming increasingly prevalent as antibiotics continue to be used indiscriminately and with waning effectiveness. It is imperative that more effective treatment modalities be advanced to adequately manage serious pulmonary infections in the clinical setting. Here we describe a highly innovative delivery and therapeutic concept, living microrobot therapeutics, for critically ill patients with severe P. aeruginosa and MRSA lung infections. The microrobot platform is consisting of Chlamydomonas reinhardtii microalgae modified with neutrophil membrane-coated and drug-loaded polymeric nanoparticles (denoted ‘algae-NP-robots’), and has unique multifold mechanisms of action. The microalgae help to improve tissue penetration and retention of the drug payload within the lungs, while the neutrophil membrane- coated nanoparticles help to shield the drug payload from biological environments, reduce immune clearance, and enable specific binding with target pathogens. Besides carrying drug payload, the neutrophil membrane- coated nanoparticles can further serve as ‘nanosponges’ that act to neutralize excessive pro-inflammatory cytokines, thus reducing the danger of cytokine storm. By combining the unique properties of these two systems, the algae-NP-robots have proven to be a capable platform for active drug delivery and excel at treating bacterial pulmonary infections. In this proposal, we describe our extensive prior published and preliminary results that strongly support the novel therapeutic concept of algae-NP-robots for the treatment of severe Gram-negative and Gram-positive pulmonary bacterial infections in ICU patients. In Aim 1, we will focus on further optimizing the algae-NP-robot formulation to maximize its therapeutic potential. In Aim 2, we seek to better understand the mechanisms by which drug-loaded algae-NP-robots can effectively clear bacterial infection using P. aeruginosa lung infection model, in which efficacy has already been demonstrated. In Aim 3, we will extend the algae-NP- robot platform for the treatment of Gram-positive pathogen (MRSA) lung infection in order to demonstrate the generalizability of the platform. Each of the Aims can be completed independently, although the information gleaned from one can be used to improve the overall approach, which can then benefit the others.

Up to $720K

Deadline: 2031-01-31

Health

Lung-on-a-Chip and Spatial Transcriptomics to Define Eicosanoid Roles in Mucosal Immunity during Influenza/Staphylococcus aureus Superinfection

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NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY/ABSTRACT Respiratory infections like the flu can lead to severe complications when bacteria invade the lungs, causing what is known as a super-infection. These infections significantly increase hospitalizations and deaths, particularly when caused by antibiotic-resistant bacteria such as Staphylococcus aureus (S. aureus). Despite medical advances, scientists do not fully understand why the immune system fails to prevent these severe infections after a viral illness. One major gap in knowledge is how mucosal immunity—the immune defense system of the lungs and airways—is disrupted following viral infections, making patients more vulnerable to bacterial super- infections. Our research focuses on a group of molecules called eicosanoids, which regulate inflammation and immune responses in the lungs. Specifically, we are studying how a subset of these molecules, called cytochrome P450 (CYP450)-derived lipids, influence immune cell behavior during bacterial super-infections. Our preliminary findings suggest that CYP450 lipids activate a protein called PPARα, which weakens the immune system's ability to fight bacteria and disrupts mucosal immunity. Mice lacking PPARα showed improved resistance to super-infections, suggesting that blocking this pathway could enhance bacterial clearance and restore lung immune defenses. Additionally, we used advanced imaging techniques to track these lipids in lung tissues and found that their distribution is linked to areas of severe infection. However, we still do not fully understand how these lipids influence immune cells at a molecular level or how they alter mucosal immunity to create conditions favorable for bacterial persistence. To address these gaps, we will use two cutting-edge approaches: 1) spatial transcriptomics, which allows us to see how different cells in the lung respond to infection at a single-cell level, and 2) a lung-on-a-chip device, which mimics human lung tissue to study real-time interactions between immune cells and bacteria. These approaches will help us understand how CYP450 lipids and PPARα signaling contribute to immune dysfunction and mucosal immunity breakdown. By uncovering the molecular mechanisms behind immune suppression and mucosal immune dysregulation during bacterial super-infection, our research has the potential to lead to new treatments. Targeting PPARα could offer a novel strategy to enhance the immune response, improve mucosal immunity, and reduce the severity of respiratory infections. This study will provide critical insights into how the body responds to lung infections and may inform future drug development to combat antibiotic-resistant bacteria.

Up to $436K

Deadline: 2028-01-31

Health

Lympnoids tissue senescence and multimodal imaging using REDCAT

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NIA - National Institute on Aging

SUMMARY Cellular senescence, a hallmark of aging, is an irreversible state of cell cycle arrest in otherwise proliferative cells. Senescence in immune organs is particularly significant in aging and disease, as immune cells circulate throughout the body and regulate the physiological functions of all major organs. Metabolic alterations play a crucial role in immunosenescence and are recognized as key mechanisms driving abnormal immune homeostasis. To better understand the distinct role of senescent cell metabolism – for example, in thymic aging and immune function – there is a pressing need for novel methods to map senescent cells and cell-type-specific metabolism within complex tissues to assess their impact on the local environment. Given the critical role of immune senescence in systemic, organism-level aging, we hypothesize that: (i) thymic epithelial cell (TEC) senescence plays a key role in driving thymic aging. (ii) abnormal metabolic dynamics, including lipid metabolism and accumulated adipocytes in aged thymi, contribute to reduced T-cell diversity, (iii) senescence and aging in the thymus lead to systemic immune function decline. To address these hypotheses, we propose to develop and deploy an integrated platform, Raman Enhanced Determination of Cell Atlas and Typing (REDCAT), for mapping single-cell metabolic activity in complex tissues and decoding the underlying transcriptional and epigenomic mechanisms. It will be applied to the study of thymic aging in wild-type, lineage-tracked, and FGF21 mouse models. Specifically, we will (1) develop REDCAT for single-cell resolution profiling of cellular senescence in complex lymphoid tissues, (2) examine senescence-associated transcriptomic mechanisms via integrating DBiT- Based spatial transcriptome sequencing, and (3) investigating dynamic phenotypic and metabolomic heterogeneity of senescent cells in thymic aging and the impact on tissue microenvironments and systemic immune function. The proposed techniques can be widely adopted by the cellular senescence and aging research community. This work will also generate a valuable data resource to unveil insights in thymic aging to advance fundamental understanding of immuno-senescence and propel translational developments in anti- senescence interventions aimed at improving immune function and overall healthspan.

Up to $701K

Deadline: 2031-01-31

Health

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