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Principal Investigator: David W Gludish
Organization: CORNELL UNIVERSITY
Fiscal Year: 2024
Award: $235,500
Funding agency: NIH Office of the Director
This project will credential two novel types of broadly applicable mouse models, which we will demonstrate in
the lung. First, we will express genes that drive disease susceptibility on one side of the lungs only, a model we
call a Unilateral Mosaic, creating disease-prone and disease resistant (internal control) lung tissue in a single
animal. The lungs serve as a proof-of-concept organ, but this Unilateral Mosaic model applies to other
asymmetric or paired organ systems such as the liver, and also to the limbs. The second type of engineered
mouse will harbor an allele that installs permanent genetic marks within niche or neighbor cells, for in vivo cellular
contact tracing in mice. Using a secreted and cell-permeant Tat-Cre protein, cells will be targeted for
recombination in vitro using co-culture and media transfer experiments, and in vivo using transplantation of
engineered Tat-Cre expressing cells into loxP-activated fluorescent reporter mice. There is an urgent need for
mouse models that reduce animal to animal variability, and increase the in vivo resolving power for tracking
cellular behavior. The models we present begin to address these challenges, and will appeal to a wide variety of
investigators that employ mice to study complex human diseases. The proposed genetic tools are designed to
address numerous categories of biological problems in living animals, but are not intrinsically confined to a
specific disease state or organ. These studies employ the well-established Cre-LoxP system for targeted in vivo
gene expression, applicable to the numerous floxed alleles in existence. This work will broadly impact the
available applications for tissue- or lineage-specific expression of genes in mice, enabling investigators to track
cells or induce gene expression in neighboring or transacting cells that contact driver cells of a given lineage.
Specific Aim 1. Generate a Unilateral Mosaic model of disease susceptibility.
a. Test whether human Ace2 excision can restrict SARS-CoV2 infection to one side of the lung.
b. Determine whether excision of Cox2/Ptgs2 unilaterally in the lungs can compartmentalize metastatic disease
after intravenous injection of metastatic mouse mammary cancer cells.
Specific Aim 2. Test and deploy a Tat-Cre allele for in vivo lineage contact tracing
We anticipate this project will yield rapid results, as we already have the ASE-Cre Unilateral Mosaic driver mice
in house and have exciting preliminary data to reveal the target organs for its asymmetric expression. We have
performed many SARS-CoV-2 viral challenge studies in human Ace2 knockin/knockout mice with a currently
approved and active animal BSL-3 program in the lab. In addition to our experience in mouse mammary tumor
models and lung developmental and cancer biology in mice, we have established collaborations to ensure the
success of syngeneic mammary tumor transplant studies. For Aim 2, we already have cloned and present
preliminary data to demonstrate the utility of the new Tat-Cre construct. In summary, we have the required
experience and existing preliminary data and materials to bring the proposed studies to a successful completion.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><ACE2><Abdomen><Abscission><Address><Affect><Alimentary Canal><Alleles><Allelomorphs><Anatomic Sites><Anatomic structures><Anatomy><Animals><Arthritis><Autoimmune Diseases><Basal Transcription Factor><Basal transcription factor genes><Biological><Biology><Blood><Blood Reticuloendothelial System><Body System><Body Tissues><Bone Tissue><Breast Cancer Model><Breast Neoplasms><Breast Tumors><Breast tumor model><Bronchopulmonary Dysplasia><COPD><COVID-19 infection><COVID-19 sequelae><COVID-19 virus><COVID-19 virus infection><COVID19 infection><COVID19 virus><CRE Recombinase><Cancer Biology><Cancers><Cardiac><Categories><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Interaction><Cell Signaling><Cell-to-Cell Interaction><Cells><Chest><Chronic Obstruction Pulmonary Disease><Chronic Obstructive Lung Disease><Chronic Obstructive Pulmonary Disease><Clinical><Co-culture><CoV-2><CoV2><Cocultivation><Coculture><Coculture Techniques><Collaborations><Communicable Disease Contact Tracing><Companions><Complement><Complement Proteins><Complex><Connective Tissue><Contact Tracing><Cre Lox technology><Cre LoxP system><Cre driver><Cre lox recombination><Cre lox recombination system><Cre lox system><Cre recombinase/LoxP technology><Cre system><Credentialing><DNA Recombination><Data><Dedications><Developmental Biology><Diathesis><Digestive Tract><Discipline><Disease><Disease Resistance><Disease susceptibility><Disorder><Dose><Embryo><Embryonic><Engineering><Enhancers><Ensure><Enterobacteria phage P1 Cre recombinase><Environment><Excision><Exhibits><Exposure to><Extirpation><Extremities><Fats><Fatty acid glycerol esters><Fibroblasts><Frequencies><Future><GI Tract><Gastrointestinal Tract><Gastrointestinal tract structure><Gene Expression><General Transcription Factor Gene><General Transcription Factors><Genes><Genetic><Genetic Recombination><Hepatitis><Human><IVIS SpectrumCT><IVIS imaging><IVIS optical imaging><IVIS spectral imaging><IVIS spectrum><IVIS system><Immunomodulation><In Vitro><In vivo analysis><Individual><Infection><Infectious Disease Contact Tracing><Inflammatory><Injections><Injury><Intracellular Communication and Signaling><Investigators><Isoforms><KI mice><KO mice><Knock-in><Knock-in Mouse><Knock-out Mice><Knockout Mice><Laboratory Animals><Left><Limb structure><Limbs><Liver><Long COVID><Long COVID-19><Long coronavirus disease><Long coronavirus disease 2019><LoxP-flanked allele><Lung><Lung Parenchyma><Lung Respiratory System><Lung Tissue><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Mammary Cancer><Mammary Neoplasms><Mammary gland><Metastasis><Metastasis to the Lung><Metastasize><Metastatic Lesion><Metastatic Mass><Metastatic Neoplasm><Metastatic Neoplasm to the Lung><Metastatic Tumor><Metastatic Tumor to the Lung><Mice><Mice Mammals><Modeling><Modern Man><Morbidity><Morbidity - disease rate><Murine><Mus><Neoplasm Metastasis><Neoplasm Transplantation><Non-Trunk><Null Mouse><Organ><Organ System><Organogenesis><Outcome><Output><Pancreas><Pancreatic><Pathology><Pathway interactions><Predisposition><Predisposition gene><Protein Isoforms><Proteins><PyMT><Receptor Protein><Recombination><Recovery><Removal><Reporter><Research Personnel><Researchers><Respiratory Disease><Respiratory System Disease><Respiratory System Disorder><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 infection><SARS-CoV-2 sequelae><SARS-CoV2><SARS-CoV2 infection><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Secondary Neoplasm><Secondary Tumor><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome related corona virus 2><Side><Signal Transduction><Signal Transduction Systems><Signaling><Spleen><Spleen Reticuloendothelial System><Stomach><Structure of parenchyma of lung><Surgical Removal><Susceptibility><Susceptibility Gene><System><Testing><Thorace><Thoracic><Thorax><Thrombus><Tissues><Transact><Transcription Factor Proto-Oncogene><Transcription factor genes><Transplantation><Validation><Viral><Viral Cancer><Viral Diseases><Viral Pneumonia><Viral Receptor><Virus Diseases><Virus Receptors><Viscera><Visceral><Work><Wuhan coronavirus><alimentary tract><angiotensin converting enzyme 2><angiotensin converting enzyme II><arthritic><autoimmune condition><autoimmune disorder><autoimmunity disease><bacteriophage P1 recombinase Cre><biologic><biological signal transduction><cancer cell><cancer metastasis><cell behavior><cellular behavior><chronic COVID><chronic COVID-19><chronic lung disease in infants><chronic lung disease in neonatal infants><chronic lung disease in neonates><chronic lung disease in newborns><chronic lung disease in prematurity><chronic lung disease in preterm infants><chronic lung disease of infancy><chronic lung disease of prematurity><chronic novel coronavirus disease 2019><chronic obstructive pulmonary disorder><complementation><coronavirus disease 2019 infection><coronavirus disease 2019 virus><coronavirus disease-19 virus><design><designing><digestive canal><disease model><disease phenotype><disorder model><experience><experiment><experimental research><experimental study><experiments><floxed><floxed allele><gastric><hCoV19><hepatic body system><hepatic organ system><human disease><human model><immune modulation><immune regulation><immunologic reactivity control><immunomodulatory><immunoregulation><immunoregulatory><in vivo><in vivo evaluation><in vivo imaging system><in vivo testing><infant chronic lung disease><infants with chronic lung disease><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><injuries><innovate><innovation><innovative><internal control><intravenous injection><knockin><knockin mice><liability to disease><long haul COVID><long haul COVID-19><long haul coronavirus disease><long haul coronavirus disease 2019><long-hauler COVID><long-hauler COVID-19><long-hauler coronavirus disease 2019><long-hauler syndrome><long-term COVID><long-term COVID-19><long-term coronavirus disease><long-term coronavirus disease 2019><longterm COVID><longterm COVID-19><longterm coronavirus disease><longterm coronavirus disease 2019><lung metastasis><lung metastatic><malignancy><mammary><mammary cancer model><mammary tumor><mammary tumor model><metastasize to the lung><model of human><mortality><mosaic><mouse model><murine model><nCoV2><neonatal chronic lung disease><neoplasm/cancer><newborn chronic lung disease><novel><paracrine><pathway><persistent COVID-19><polyoma middle tumor antigen><post COVID syndrome><post COVID-19 syndrome><post acute COVID syndrome><post acute COVID-19><post acute COVID-19 syndrome><post acute SARS-CoV-2><post acute coronavirus disease 2019><post acute coronavirus disease 2019 syndrome><post acute coronavirus disease syndrome><post acute severe acute respiratory syndrome coronavirus 2><post coronavirus disease 2019 syndrome><post coronavirus disease syndrome><post-acute phases of COVID-19><post-prematurity respiratory disease><predisposing gene><preterm infants with chronic lung disease><programs><prolonged COVID-19 symptoms><promoter><promotor><pulmonary><pulmonary metastasis><pulmonary 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