Document text
Principal Investigator: Jacob Brenner
Organization: BIOSPUTNIK LLC
Fiscal Year: 2023
Award: $301,016
Funding agency: National Human Genome Research Institute
ABSTRACT
Lymphangioleiomyomatosis (LAM) is a Tuberous sclerosis-related disorder. Both occur due to an
inherited or sporadic mutation in either the TSC1 or TSC2 gene, which function as negative
regulators of the mTOR pathway. Uncontrolled mTORC1 activity leads to the neoplastic
proliferation of abnormal smooth muscle cells (LAM cells) in the lungs, progressive shortness of
breath, recurrent pneumothoraxes, and loss of pulmonary tissue structure and function primarily
in women. The first and only FDA-approved treatment for LAM is the immunosuppressant
sirolimus, marketed since 2015 by Pfizer. It is the current standard-of-care and acts by inhibiting
mTORC1. Sirolimus has several clinical disadvantages, including a considerable number of non-
responders, severe adverse events due to its immunosuppressive properties and pregnancy
category C, limiting its use in women of childbearing age. Thus, there is a high unmet medical need
to develop alternative and safer treatment options for LAM and TS. We have identified treatment
with miRNA302b mimics as a potential novel therapy for LAM/TS. Using a murine lung injury
model, our collaborator Hao Shen was able to show that non-targeted treatment with miRNA302b
mimics as neutral lipid emulsion improved lung function, host recovery, and alveolar epithelial
regeneration mice. We also demonstrated a “stalled” AT2-AT1 transdifferentiation state in human
LAM, suggesting that impaired AT2 fitness may contribute to loss of alveolar structure. Our goal
for this grant is the investigation of pulmonary-epithelium targeted miRNA302b mimic lipid
nanoparticles (LNP) efficacy for the treatment of LAM by enhancing AT2 cell regeneration. Aim 1
is composed of in vitro characterization of the miRNA-302b mimic lipid nanoparticle and
investigation of several targeting strategies in vivo. Our PI Dr. Jake Brenner will design and
construct the nanoparticles in his bioengineering lab. We will perform 3D organoid experiments
with human and mouse AT2 cells in the presence of the LNP[miR302b] followed by an in vivo study
comparing different targeting strategies to reach the desired tissue and cell type using a transgenic
model of LAM, developed by our other PI Prof. Vera Krymskaya. Using the most effective targeting
strategy, Aim 2 will be an in vivo proof-of-concept study using that same transgenic murine model
pre and post-pregnancy to answer the question whether our treatment will prevent airspace
enlargement and lead to lung recovery. We will observe mice for 4 and 8 weeks after intratracheal
LNP[miR302b] administration. Endpoints will include lung function, BALF analysis, qPCR, and
histological lung sections stained for the disease related markers and cell types. All collaborators
are experts in their respective field and thus well equipped to successfully complete this project,
bringing together a number of unique and innovative aspects to treat this devastating rare disease.
Terms: <3-D><3-Dimensional><3D><Acceleration><Adverse effects><Affect><Age><Alveolar><Anti-Oncogenes><Antibodies><Antioncogenes><Biomedical Engineering><Body Tissues><Bourneville Disease><Bourneville Phakomatosis><Bourneville syndrome><Bourneville-Brissaud disease><Bourneville-Pringle syndrome><CD47><CD47 Antigen><CD47 Glycoprotein><CD47 gene><Cancer Suppressor Genes><Categories><Cell Body><Cell Communication and Signaling><Cell Culture Techniques><Cell Signaling><Cells><Clinical><Cyst><DNA><Data><Deoxyribonucleic Acid><Disadvantaged><Disease><Disease Progression><Disorder><Drug Delivery><Drug Delivery Systems><Drugs><Emerogenes><Emulsions><Ensure><Epiloia><Epithelial Cells><Epithelium><Equilibrium><FDA approved><FK506 Binding Protein 12-Rapamycin Associated Protein 1><FKBP12 Rapamycin Complex Associated Protein 1><FRAP1><FRAP1 gene><FRAP2><Family><Female><Female of child bearing age><Female of childbearing age><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Formulation><Genetic><Genetic Alteration><Genetic Change><Genetic defect><Gestation><Goals><Grant><Hamartin><Hereditary><Heterozygote><Histologic><Histologically><Human><Human Characteristics><Human Nature><Immune response><Immunological response><Immunosuppressants><Immunosuppressive Agents><Immunosuppressive drug><Immunosuppressive treatment><Impairment><In Vitro><Inherited><Integrin-Associated Protein><Intercept><Intracellular Communication and Signaling><Intravenous><Investigation><Involuntary Muscle><Leiomyocyte><Lesion><Life><Lipids><Liposomal><Liposomes><Lung><Lung Alveolar Epithelia><Lung Function Tests><Lung Lymphangioleiomyomatosis><Lung Respiratory System><Lung damage><Lung diseases><Lymphangioleiomyomatosis><Lymphangiomyomatosis><MER6><Marketing><Measures><Mechanistic Target of Rapamycin><Medical><Medication><Mesenchymas><Mesenchyme><Mice><Mice Mammals><Micro RNA><MicroRNAs><Microscopic><Modeling><Modern Man><Murine><Mus><Mutation><Nanotechnology><Natural regeneration><Non-Polyadenylated RNA><Onco-Suppressor Genes><Oncogenes-Tumor Suppressors><Organoids><Orphan Disease><Pathway interactions><Patients><Persons><Pharmaceutic Preparations><Pharmaceutical Preparations><Pneumothorax><Pregnancy><Prevention><Pringle disease><Progenitor Cells><Proliferating><Property><Pulmonary Diseases><Pulmonary Disorder><Pulmonary function tests><Qualifying><RAFT1><RNA><RNA Gene Products><Rapamune><Rapamycin><Rare Diseases><Rare Disorder><Recessive Oncogenes><Recovery><Recurrence><Recurrent><Regeneration><Research><Ribonucleic Acid><Route><SDZ RAD><Sampling><Scientist><Serious Adverse Event><Severe Adverse Event><Shortness of Breath><Signal Transduction><Signal Transduction Systems><Signaling><Sirolimus><Smooth Muscle><Smooth Muscle Cells><Smooth Muscle Myocytes><Smooth Muscle Tissue Cell><Staining method><Stains><Structure><Surface><Surface Antigen Identified by Monoclonal Antibody 1D8><System><TSC1><TSC1 gene><TSC2><TSC2 gene><TSC4><TSC4 Gene><Testing><Tissues><Transgenic Mice><Transgenic Model><Transgenic Organisms><Treatment Efficacy><Tuberin><Tuberous Sclerosis><Tumor Suppressing Genes><Tumor Suppressor Genes><United States><Woman><adenoma sebaceum><after pregnancy><age dependent><age related><ages><alveolar epithelium><balance><balance function><bio-engineered><bio-engineers><bioengineering><biological engineering><biological signal transduction><cell culture><cell cultures><cell regeneration><cell type><cellular regeneration><cerebral sclerosis><clinical development><cohort><conditional knock-out><conditional knockout><design><design and construct><design and construction><designing><disease of the lung><disorder of the lung><drug/agent><epiploia><epithelium regeneration><everolimus><experiment><experimental research><experimental study><experiments><fetal><fitness><flow cytophotometry><gain of function><genome mutation><hereditary multiple system hamartomatosis><heterozygosity><host response><immune suppressive agent><immune suppressor><immune system response><immunoresponse><immunosuppressive substance><immunosuppressor><improved><in vivo><innovate><innovation><innovative><intervention efficacy><lipid based nanoparticle><lipid nanoparticle><lung development><lung disorder><lung function><lung function decline><lung injury><lung lesion><mTOR><mammalian target of rapamycin><miRNA><miRNA delivery><miRNAs><microRNA delivery><mouse model><murine model><nano particle><nano tech><nano technology><nano-sized particle><nano-technological><nanocarrier><nanoparticle><nanosized particle><nanotech><nanotechnological><nanovessel><neoplastic><neurinomatosis centralis><neuromatosis universalis><neurospongioblastosis diffusa><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><oncosuppressor gene><orphan disorder><pathway><phacomatosis><pneumothorax disorder><post pregnancy><pregnant><prepregnancy><prevent><preventing><pulmonary><pulmonary damage><pulmonary function><pulmonary function decline><pulmonary injury><pulmonary lesion><pulmonary lymphangioleiomyomatosis><pulmonary tissue damage><pulmonary tissue injury><rare genetic disease><rare genetic disorder><regenerate><regenerate epithelium><regeneration based therapy><regeneration therapy><regenerative therapeutics><regenerative therapy><repair><repaired><restoration><sclerosis tuberosa><serious adverse experience><serious adverse reaction><sex><site targeted delivery><small molecule><spongioblastosis circumscripta><standard of care><stem cells><success><targeted delivery><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic efficacy><therapy efficacy><three dimensional><tissue repair><transdifferentiation><transgenic><transgenic trait><tuberose sclerosis><tuberous sclerosis 1><tuberous sclerosis complex><women of child bearing age><women of childbearing age>