Research Portfolio

It is amazing how far LAM research has come in such a short period of time. Many researchers who have received funding from The LAM Foundation have helped to make this disease a research priority. LAM and TSC researchers have identified a wealth of potential molecular targets and experimental therapies that may be appropriate for testing in clinical trials. Many of these drugs are FDA-approved or in development for other indications.

The LAM Foundation has committed over $11 million to LAM research to support 119 peer-reviewed grants and other research projects for the study of lymphangioleiomyomatosis (LAM). Scientists funded by The LAM Foundation reported numerous breakthroughs, resulting in the first-ever LAM treatment trial to test a drug called sirolimus, or rapamycin, which proved to be an effective treatment for LAM (results published in the April 2011 issue of the New England Journal of Medicine (NEJM). Additional clinical trials are underway as LAM Foundation-funded researchers continue to work with urgency in search of more safe and effective treatments.

2015 Award Winnersplus

Established Investigator Awards - $150,000 over three years
Grant Award Winners
I. Caroline Le Poole, PhD
Loyola University
Chicago

T Cell Receptor Transduced T Cells Supported by Anti-PD-1 to Treat LAM

LAM cells share some features with the cells that make pigment in the skin. The molecules that are responsible for pigmentation can be misread as foreign particles by the immune system in patients that develop the autoimmune disease vitiligo. We thus isolated immune cells from vitiligo patient skin and identified the recognition molecule on the surface of T cells that allows these immune cells to recognize gp100. Expression of gp100 is also a hallmark of LAM, and HMB45 staining can be used to detect it and confirm a LAM diagnosis in patients. We now have the gp100-reactive T cell receptor in hand so that it can be introduced into patient T cells to be redirected towards LAM tissue. The gp100-reactive T cells may become exhausted before the diseased tissue is effectively eliminated by the redirected T cells. In fact, we have seen that our LAM cell line is equipped with the tools to exhaust reactive T cells. We thus want to test the anti-PD1 reagent, recently approved for use in melanoma and in lung cancer, for its value in the treatment of LAM, to prevent exhaustion in our redirected T cells. To put this combination therapy to the test, we will make use of a preclinical model of LAM using a human LAM cell line and redirected human T cells. This live model allows for the outgrowth of traceable human LAM cells and their interaction with T cells in presence of the anti-PD-1 reagent. The outcome of these studies may serve as the basis for clinical studies in (wo)man.

Grant Award Winners
Kuniaki Seyama, MD, PhD
Jutendo University
Tokyo, Japan

Exploring Pathobiology of LAM Through Isolation of Cellular Components of LAM Lesions, Their Characterizations and the Reconstruction of LAM Lesions by Patient-Derived LAM Cells and Lymphatic Endothelial Cells

The prevailing LAM models at present are -knockout mice or a mouse model explanting -null cell line or human -null angiomyolipoma cells. However, these have some faults since they unfortunately lack lymphatic endothelial cells (LEC), an important component of LAM lesions. In addition, no human LAM cells with mutations have been isolated from LAM patients. Therefore, the pathobiology of LAM would have not been fully disclosed by the existing models. We are going to establish a flow cytometry (FACS)-based method by which that HMB45-positive and mutation-harboring LAM cells can be isolated from human LAM lung tissues. In addition, other cellular components of LAM lesions, e.g. LEC and mesenchymal cells, are simultaneously obtained. We would like to establish the culture technique to expand these cellular components , and then reconstitute LAM lesions in a 3-dimedional gel. We expect that this reconstitute would work as a new “ LAM model” to explore what kind of cell-cell interactions work there and what the key mechanism for LAM cells survive.

Pilot Awards -$25,000 for one year
Grant Award Winners
Michael Borchers, PhD
University of Cincinnati
College of Medicine
Cincinnati

NKG2D Receptor-Ligand Interactions in LAM Pathogenesis

Lymphangioleiomyomatosis (LAM) is a rare lung disease of women in which smooth muscle cells from an unknown source proliferate and infiltrate the lung. Proliferating smooth muscle cells lead to destruction of lung tissue, leading to respiratory failure. Typically, abnormally proliferating cells are recognized by immune cells through expression of surface markers that target them for elimination. It is important to understand how LAM cells evade this immune detection. Recognition of abnormal cells by cytotoxic lymphocytes is mediated in part through the NKG2D receptor that recognizes ligands on abnormal cells. NKG2D ligand expression is highly restricted and absent on healthy cells, but abundant on the surface of tumor cells. The importance of NKG2D–mediated control of abnormal cell proliferation is established. However, recent studies demonstrate that the elaboration of soluble NKG2D ligands suppress tumor immunity by acting as a sink and downregulating NKG2D expression and function. This proposal will test the possibility that soluble ligands for the NKG2D receptor are useful biomarkers for disease progression. Further, we will examine the function of the NKG2D receptor in LAM patients to gain insight into disease progression. Several therapeutics aimed at modulating NK cell receptors are in development for chronic diseases, autoimmune diseases, and cancer. Our hope is that these novel agents will provide exciting therapeutic options for LAM patients in the near future.

Grant Award Winners
Joshua Fessel, MD, PhD
Vanderbilt University
Nashville

Use of 18F-Glutamine PET in the Diagnosis of LAM and Other Pulmonary Diseases

Diagnosing many lung diseases, including LAM, can be challenging. Though sometimes a disease can be identified by a combination of physical exam findings, imaging findings (like X-rays or CT scans), and blood tests, often a definitive diagnosis requires a lung biopsy. This can be difficult to accomplish safely, and even under the best of circumstance, there is always some degree of risk and inconvenience involved. This project hopes to improve the use of special imaging techniques so that LAM and other lung diseases can be more confidently diagnosed without lung biopsies, and so that responses to existing and new treatments can be monitored more easily.

2014 Award Winnersplus

Established Investigator Awards - $150,000 over three years
Grant Award Winners
Lisa Young, MD
Vanderbilt University
Nashville

Lymphangiogenesis and Estrogenic Milieu as LAM Biomarkers

Lymphangioleiomyomatosis (LAM) is a progressive lung disease that occurs essentially only in women. Vascular endothelial growth factors (VEGF) promote the growth of blood/lymphatic vessels. Serum VEGF-D is a protein that is elevated in the blood of ~70% of women with LAM and can be used for LAM diagnosis. In addition, higher VEGF-D levels are associated with more severe lung disease, and blood levels correlate with response to sirolimus therapy. However, VEGF-D levels are normal in ~30% of LAM patients, and there remains significant unmet need for additional biomarkers to aid in diagnosis, prognosis, and assessment of treatment response. Further, the reason why LAM occurs only in women remains uncertain, as well as the factors that lead to onset of LAM in women with Tuberous Sclerosis Complex (TSC). This research project is designed to determine whether VEGF-D levels are elevated in women with TSC who are at risk for LAM, and to examine whether other related markers are elevated in the blood of women who have normal VEGF-D levels. These studies will also examine measures of estrogen activity, to understand whether estrogen-related compounds are additional biomarkers for LAM that may relate to disease onset and progression. Overall, these studies will improve understanding of the early contributors to LAM onset and disease progression, and identify additional biomarkers that can be used to diagnose, predict prognosis, and monitor patients with LAM.

Grant Award Winners
Andrey Parkhitko, PhD
Harvard Medical School
Boston

A Cross-Species Approach to the Discovery of Genes Accelerating TSC/LAM Tumor Growth

The goal of this research project is to test the relevance of the additional transcriptional alterations in Tuberous sclerosis complex (TSC) tumors to its pathogenesis. TSC, a tumor syndrome associated with various tissues that include the brain, skin, kidneys, heart, and lungs, affects an estimated 1 in 6,000 to 10,000 births. In addition, varied neurological and cognitive deficits are very common and represent the most severe features of TSC. Currently, clinical trials of mTORC1 inhibitors (Sirolimus, Everolimus) are effective at reducing the size of SEGA and renal AMLs, as well as stabilizing lung function in women with LAM. After withdrawal, however, renal AMLs regrow and lung function decline resumes. Our research will allow the identification of additional pathways that synergize with mTORC1 and provide new strategies for the cure of TSC diseases.

Pilot Awards -$25,000 for one year
Grant Award Winners
Raymond Yeung, MD
University of Washington
Seattle

Serum Metabolites in LAM

The diagnosis of LAM is often delayed and requires invasive biopsy. Blood tests such as VEGF-D level may one day simplify this process. Biomarkers are also useful in detecting disease burden and may indicate response to therapy. Hence, biomarkers may enable one to determine if a certain treatment is likely to produce a response or not, and if not, to seek alternatives and to lessen undue toxicities. In this proposal, we seek to discover novel biomarkers using state-of-the-art techniques in detecting changes in metabolite levels in the blood. This pilot award will enable us to screen a limited number of samples and come up with a handful of candidate metabolites that will form the basis of larger validation studies in the future.

Grant Award Winners
Debbie Clements, PhD
Nottingham University
United Kingdom

Investigating the Cross Talk Between LAM Cells and recruited Stromal Fibroblasts

LAM cells grow in the lungs and cause the lung tissue to break down, resulting in respiratory failure, but we do not know what causes this to happen. Our hypothesis is that the nodules that form in the lungs of patients with LAM contain different cell types, some of which have a mutation in TSC2, and some of which do not. We think the cells which do not carry the mutation are a normal cell type called a fibroblast. We believe that it is the interaction between these cells that is so destructive to the lungs, because high levels of damaging proteases are generated.

We would like to test our hypothesis by looking very closely at changes that occur in fibroblasts after they have been in contact with LAM cells. The cells may become more migratory, grow faster, or produce more proteases. We are able to look at changes in the expression of all the genes of the fibroblasts at once using a microarray. We can then see which pathways are most affected by the interaction between the cells and identify underlying mechanisms that will be targets for new and effective therapies.

Once we have identified altered pathways we can investigate them in a model of LAM, including both fibroblasts and cells with a mutation in TSC2, which we can grow in a tissue culture dish. We can target these pathways using existing drugs and determine whether this disrupts the formation or survival of our co-culture LAM nodules, and in this way test many drugs which may prove to be novel therapies for LAM.

Grant Award Winners
Khalid Almoosa, MD
University of Texas
Health Science Center

Does Variability of Care Exist Among LAM Specialty Clinics?

Variability in healthcare is a common problem that can affect the quality of care provided to patients with complex diseases. Lymphangioleiomyomatosis (LAM) is a rare and complex lung disease that affects women and results in significant limitations in their health and life activities. The LAM Foundation has helped establish specialty clinics for LAM patients whose goal is to provide the best and most state-of-the-art care possible. The objective of this application is to determine the presence of unwarranted variability in the LAM clinics that is not due to patients needs or preferences and reduce it, so that the optimal quality of care is provided to these patients at all times. The successful completion of this project will further our understanding of how these growing LAM clinics operate, including their strengths, challenges and patient experiences.

2013 Award Winnersplus

Pilot Clinical Trial Award
Grant Award Winners
Vera Krymskaya, PhD, MBA
University of Pennsylvannia
Philadelphia

The Safety of Simvastatin in Patients with Sporadic Pulmonary Lymphangioleiomyomatosis (LAM) and LAM Associated with Tuberous Sclerosis Complex (The SOS Trial)

Pulmonary LAM, which can be sporadic (LAM-S) or associated with tuberous sclerosis (LAM-TS), is a rare lung disease affecting predominantly women of childbearing age that manifests by neoplastic growth of atypical smooth muscle-like LAM cells, lung cyst formation, obstruction of lymphatics, and spontaneous pneumothoraces (1). Current treatment with rapamycin analogs, which are not FDA-approved for treatment of LAM-S and LAM-TS, but are commonly used based upon published reports, stabilize lung function but does not prevent further disease progression (1, 2). The rapalogs are cytostatic agents and cessation of treatment resumes decline in lung function. Because of this limitation new combinational therapy targeting LAM cell death is needed. Preclinical evidence demonstrates that simvastatin acts as a pro-apoptotic agent in in vitro and in vivo studies in cells and tumor lesions deficient for TSC2, inactivation mutation of which cause LAM and TS (3, 4). HMG-CoA reductase inhibitors (statins) are very safe, highly effective therapies used by millions of people (5, 6). Simvastatin decreases cholesterol, reduces cancer incidence, stabilizes the endothelial cell layer and decreases inflammation. In preclinical studies the combination of rapamycin and simvastatin inhibited xenograft tumor growth and completely blocked the recurrence of xenograft tumors after treatment withdrawal (4). In mouse model of LAM, treatment with rapamycin and simvastatin prevented both growth of TSC2-null lesions and lung destruction (3). These findings have fundamental translational significance for combinatorial therapeutic strategies to induce the death of LAM cells, potentially obviating the need for continuous, life-long suppressive therapies. We have designed a Phase II, open label, safety trial to initiate the study of this potentially useful intervention. We propose to conduct a safety trial of simvastatin treatment of patients with LAM-S and LAM-TS. We aim to determine the safety of simvastatin in combination with sirolimus or everolimus over 4 months.

Designated Award
Grant Award Winners
David Kwiatkowski, MD, PhD
Brigham and Women’s Hospital
Boston

Genomewide Association Studies (GWAS)

Genome wide association studies (GWAS) are a type of study commonly used by medical genetic researchers to try to identify genes and their variants that are associated with, or in some way predispose to, human disease. They have been performed for almost every common disease you can think of, including diabetes, cancer, and heart disease. A GWAS has not been performed in LAM previously. We suspect that genetic risk may be significant in LAM given the strong association of LAM development with female sex and the consistent occurrence of TSC2 gene mutations in LAM cells. One great thing about GWAS is that they are unbiased. This means that the entire genome is scrutinized, so that any gene anywhere that has a role in LAM development may be discovered. It is well-known that many genes have been identified in this manner in other diseases by GWAS which were completely unexpected. Therefore, there is the possibility that something completely different from what we know about LAM will be discovered from GWAS. Therefore, we propose to perform a GWAS of LAM with appropriate follow-up studies.

Pilot Awards -$25,000 for one year
Grant Award Winners
Gina Lee, PhD
Harvard Medical School
Boston

Mechanism of Tumorigenesis by mTORC1-Medicated mRNA Alternative Splicing

Cells can make different types of proteins from a single gene by cutting, removing, and rejoining part of the gene. This process, called alternative splicing, normally occurs to make specific protein types required for each organ and developmental time. Cancer research has mostly focused on investigating gene expression or protein quality control. However, recent large-scale screen technologies have found that alternative splicing is remarkably changed in tumors. For example, cancer cells express an embryonic type of a certain metabolic protein to provide growth advantage. In breast cancer patients resistant to chemotherapy, aberrantly spliced oncogene was identified to confer drug resistance. I discovered that alternative splicing of specific genes is changed in LAM cells. Moreover, I found that the activity of splicing regulating factors is also dysregulated in LAM cells. I will perform in-depth mechanistic studies to identify signaling processes controlling alternative splicing and investigate the effect of altered splicing in LAM pathogenesis. Successful completion of the proposed work will demonstrate that the alternative splicing process is likely to be a rich source of novel drug targets and diagnostic markers, which will lead to development of more selective and effective therapeutic interventions for LAM.

Grant Award Winners
Chenggang Li, PhD
Brigham and Women’s Hospital
Boston

Estradiol and mTORC2 Orchestrate to Enhance Prostaglandin Biosynthesis and Tumorigenesis in LAM

Lymphangioleiomyomatosis (LAM), the pulmonary manifestation of tuberous sclerosis complex (TSC), is a devastating disease affecting women, often leading to end-stage lung disease during or soon after pregnancy. The pathogenesis of LAM is very unusual: LAM cells are histologically-benign smooth muscle cells carrying TSC2 mutations that are believed to transfer or metastasize to the lungs, where they cause lung degeneration. The only proven treatment for LAM is lung transplantation, which carries significant one-year mortality and after which LAM can recur in the transplanted lungs. The reasons that LAM affects women almost exclusively are not yet clearly defined, and animal models that mimic the metastatic behavior of LAM cells have not been previously developed. Our animal model provides in vivo evidence that estrogen promotes the survival and metastasis of Tsc2-null cells and increased expression of COX-2, which allows us to investigate the possibility that targeting COX-2 pathways may have a major role in the treatment of TSC and LAM. In addition, this model will allow other novel agents and/or combinations of existing agents to be tested in preclinical stage. This will further facilitate the eventual development of effective therapies for TSC and LAM, which are urgently needed.

Grant Award Winners
Hilaire Lam, PhD
Brigham and Women’s Hospital
Boston

MicroRNA in LAM: Therapeutic Targets and Biomarkers

Lymphagioleiomyomatosis (LAM) is a devastating disease of women that is associated with lung destruction and progressive lung function decline. LAM is caused by mutations in TSC1 and 2, resulting in hyperactive mammalian Target of Rapamycin (mTORC1), which leads to excessive cell growth and proliferation. Rapamycin and related “rapalogs” inhibit mTORC1 and promote tumor regression; however, tumors regrow upon treatment cessation suggesting that rapalogs prevent LAM cell proliferation, but do not induce specific LAM cell death. To identify novel therapies that promote cell death in TSC-deficient cells, we sought to determine whether dysregulated expression of microRNA (miRNA or miR) contributes to TSC pathogenesis and rapamycin-resistance. miRNA are small molecules that can target multiple mRNA transcripts, negatively regulating the formation of proteins from genes. In recently published experiments using TSC2-deficient cells we identified rapamycindependent miRNA “Rapa-miRs”. Amongst the most highly expressed Rapa-miRs we identified pro-tumorigenic miRNA, “onco-miRs,” including miR-21, miR-221 and miR-29b. We hypothesize that targeting these pro-survival miRNAs will enhance rapamycin effectiveness. To explore this hypothesis we will investigate the regulation of miRNA by rapamycin, identify target genes regulated by rapa-miRs, determine the in vitro and in vivo effects of rapa-miR knockdown, and measure TSCLAM miRNA biomarkers in patient plasma versus healthy patient controls. These studies will identify novel therapeutics and potential biomarkers, which may have a profound impact on the clinical diagnosis and treatment of LAM patients.

To see all grants awarded by The LAM Foundation, download our entire research portfolio.