Lymphangioleiomyomatosis (LAM), characterized by nodular proliferation of abnormal smooth muscle- like cells (LAM cells) and pulmonary cysts, is a significant cause of morbidity and mortality in women with Tuberous Sclerosis Complex (TSC) or with the sporadic form of LAM. LAM is associated with genetic inactivation of TSC1 or TSC2. Critical knowledge gaps include the heterogeneity of cells within LAM nodules, the cellular origin of TSC2-deficient LAM cells, and mechanisms underlying the female predominance of LAM. Early developmental deletion of Tsc2 (E6.5-E14.5) in lung mesenchyme resulted in nodular pulmonary lesions in adulthood, predominantly in female mice. These proliferative nodules are composed of cells with loss of Tsc2 as well as wild-type cells, resembling human LAM nodules. The nodules contain cells expressing smooth muscle and melanocyte lineage proteins, hallmarks of human LAM. Later developmental deletion of Tsc2 (E13.5-P1), sparing the pulmonary vasculature, does not result in pulmonary nodular lesions in adulthood. Metabolic and mitochondrial defects are present in Tsc2-null mesenchymal cells derived from the pulmonary nodules, compared with wild-type controls. Estrogen stimulates proliferation of these Tsc2-null cells, but not wild-type cells, and upregulates mitochondrially-derived reactive oxygen species. These and other preliminary data lead to our central hypothesis: hyperactivation of mTORC1 in subsets of lung mesenchymal cells leads to estrogen-dependent proliferation of nodular-like lesions, associated with metabolic and mitochondrial dysfunction, recruitment of extrapulmonary cells and lung destruction.
Two specific aims are proposed: 1) To dissect the molecular and cellular mechanisms of LAM using a novel mouse model with spontaneous LAM-like lung lesions. The role of hyperactivated mTORC1 in the pulmonary LAM-like nodules will be examined using both pharmacologic and genetic approaches. The impact of Tsc2 deficiency on cellular metabolism and mitochondrial function will be addressed for the first time in lung mesenchyme-derived cells as a potential mechanism for the abnormal cell growth. The origin of LAM cells and the mechanisms of recruitment of extrapulmonary mesenchymal cells to LAM-like nodules will be examined using both parabiosis and allograft models. 2) To determine the specific role of gender in contributing to the formation and progression of LAM-like nodules in mice with lung mesenchymal Tsc2 deletion. The roles of estrogen in the pulmonary LAM-like lesions will be investigated in vivo by altering estrogen activity or simultaneous deletion of ER?/Tsc2 or ER?/Tsc2 and in vitro by metabolic profiling, RNA-seq, ChIP-seq in Tsc2-null vs. wild-type cells. This project will have high scientific and clinical impact by filling knowledge gaps with translational impact for women with LAM and providing a new preclinical therapeutic model.

Public Health Relevance

This project is focused on the pathogenic mechanisms through which mesenchymal loss of Tsc2 causes pulmonary lymphangioleiomyomatosis (LAM)-like nodular lesions with female predominance. We expect our results to fill several key knowledge gaps in the LAM research field, with high clinical impact for women with Tuberous Sclerosis Complex (TSC) and/or LAM. Finally, the mouse model studied in this project appears to represent the first genetically engineered mouse with spontaneous LAM-like lung pathology, providing a novel platform for preclinical therapeutic investigation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL146541-02
Application #
9900580
Study Section
Lung Injury, Repair, and Remodeling Study Section (LIRR)
Program Officer
Vuga, Louis J
Project Start
2019-04-01
Project End
2023-03-31
Budget Start
2020-04-01
Budget End
2021-03-31
Support Year
2
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Children's Hospital of Los Angeles
Department
Type
DUNS #
052277936
City
Los Angeles
State
CA
Country
United States
Zip Code
90027