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Real-time monitoring of tumorigenesis, dissemination, & drug response in a preclinical model of lymphangioleiomyomatosis/tuberous sclerosis complex.

Liu F, Lunsford EP, Tong J, Ashitate Y, Gibbs SL, Yu J, Choi HS, Henske EP, Frangioni JV - PLoS ONE (2012)

Bottom Line: Surprisingly, TSC2-deficient cells administered intratracheally resulted in rapid dissemination to lymph node basins throughout the body, and histopathological changes in the lung consistent with LAM.Estrogen was found to be permissive for tumor growth and dissemination.We generated homogeneous NIS/GFP co-expressing TSC2-deficient, patient-derived cells that can proliferate and migrate in vivo after intratracheal instillation.

View Article: PubMed Central - PubMed

Affiliation: Division of Hematology/Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT

Background: TSC2-deficient cells can proliferate in the lungs, kidneys, and other organs causing devastating progressive multisystem disorders such as lymphangioleiomyomatosis (LAM) and tuberous sclerosis complex (TSC). Preclinical models utilizing LAM patient-derived cells have been difficult to establish. We developed a novel animal model system to study the molecular mechanisms of TSC/LAM pathogenesis and tumorigenesis and provide a platform for drug testing.

Methods and findings: TSC2-deficient human cells, derived from the angiomyolipoma of a LAM patient, were engineered to co-express both sodium-iodide symporter (NIS) and green fluorescent protein (GFP). Cells were inoculated intraparenchymally, intravenously, or intratracheally into athymic NCr nu/nu mice and cells were tracked and quantified using single photon emission computed tomography (SPECT) and computed tomography (CT). Surprisingly, TSC2-deficient cells administered intratracheally resulted in rapid dissemination to lymph node basins throughout the body, and histopathological changes in the lung consistent with LAM. Estrogen was found to be permissive for tumor growth and dissemination. Rapamycin inhibited tumor growth, but tumors regrew after the drug treatment was withdrawn.

Conclusions: We generated homogeneous NIS/GFP co-expressing TSC2-deficient, patient-derived cells that can proliferate and migrate in vivo after intratracheal instillation. Although the animal model we describe has some limitations, we demonstrate that systemic tumors formed from TSC2-deficient cells can be monitored and quantified noninvasively over time using SPECT/CT, thus providing a much needed model system for in vivo drug testing and mechanistic studies of TSC2-deficient cells and their related clinical syndromes.

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The effect of sex and exogenous estrogen on TSC2-deficient tumor growth.Female (left) or male (right) mice were implanted with 0.18 mg 17ß-estradiol pellets or control placebo pellets subcutaneously. Ten days later, 621-327 cells were administered intratracheally and tumors quantified every other week by SPECT/CT. Shown are the results after 4 weeks of hormone treatment. T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors. Scale bars  = 1 cm. +/−  = 0.3–1.4; +  = 1.5 - 2.5; ++  = 2.6–3.7; +++  =  ≥3.8%ID/mm3 (×10−5).
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pone-0038589-g005: The effect of sex and exogenous estrogen on TSC2-deficient tumor growth.Female (left) or male (right) mice were implanted with 0.18 mg 17ß-estradiol pellets or control placebo pellets subcutaneously. Ten days later, 621-327 cells were administered intratracheally and tumors quantified every other week by SPECT/CT. Shown are the results after 4 weeks of hormone treatment. T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors. Scale bars  = 1 cm. +/−  = 0.3–1.4; +  = 1.5 - 2.5; ++  = 2.6–3.7; +++  =  ≥3.8%ID/mm3 (×10−5).

Mentions: LAM occurs almost exclusively in women, with onset usually during the childbearing years, which suggests that estrogen may play a role in disease progression. To measure the effect of estrogen, we implanted male and female mice with estradiol pellets or placebo pellets 7 to 10 days before inoculation of tumor TSC2-deficient cells. When assessed at 4 weeks, the 621-327 cells had a much lower initiation and growth rate in male mice, and lower overall radiotracer accumulation, which was partially corrected by estradiol supplementation (Figure 5). Significant radiotracer uptake was seen in female mice implanted with either estradiol or placebo, suggesting that endogenous levels of estrogen in female mice are sufficient for tumor initiation and proliferation.


Real-time monitoring of tumorigenesis, dissemination, & drug response in a preclinical model of lymphangioleiomyomatosis/tuberous sclerosis complex.

Liu F, Lunsford EP, Tong J, Ashitate Y, Gibbs SL, Yu J, Choi HS, Henske EP, Frangioni JV - PLoS ONE (2012)

The effect of sex and exogenous estrogen on TSC2-deficient tumor growth.Female (left) or male (right) mice were implanted with 0.18 mg 17ß-estradiol pellets or control placebo pellets subcutaneously. Ten days later, 621-327 cells were administered intratracheally and tumors quantified every other week by SPECT/CT. Shown are the results after 4 weeks of hormone treatment. T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors. Scale bars  = 1 cm. +/−  = 0.3–1.4; +  = 1.5 - 2.5; ++  = 2.6–3.7; +++  =  ≥3.8%ID/mm3 (×10−5).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3376142&req=5

pone-0038589-g005: The effect of sex and exogenous estrogen on TSC2-deficient tumor growth.Female (left) or male (right) mice were implanted with 0.18 mg 17ß-estradiol pellets or control placebo pellets subcutaneously. Ten days later, 621-327 cells were administered intratracheally and tumors quantified every other week by SPECT/CT. Shown are the results after 4 weeks of hormone treatment. T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors. Scale bars  = 1 cm. +/−  = 0.3–1.4; +  = 1.5 - 2.5; ++  = 2.6–3.7; +++  =  ≥3.8%ID/mm3 (×10−5).
Mentions: LAM occurs almost exclusively in women, with onset usually during the childbearing years, which suggests that estrogen may play a role in disease progression. To measure the effect of estrogen, we implanted male and female mice with estradiol pellets or placebo pellets 7 to 10 days before inoculation of tumor TSC2-deficient cells. When assessed at 4 weeks, the 621-327 cells had a much lower initiation and growth rate in male mice, and lower overall radiotracer accumulation, which was partially corrected by estradiol supplementation (Figure 5). Significant radiotracer uptake was seen in female mice implanted with either estradiol or placebo, suggesting that endogenous levels of estrogen in female mice are sufficient for tumor initiation and proliferation.

Bottom Line: Surprisingly, TSC2-deficient cells administered intratracheally resulted in rapid dissemination to lymph node basins throughout the body, and histopathological changes in the lung consistent with LAM.Estrogen was found to be permissive for tumor growth and dissemination.We generated homogeneous NIS/GFP co-expressing TSC2-deficient, patient-derived cells that can proliferate and migrate in vivo after intratracheal instillation.

View Article: PubMed Central - PubMed

Affiliation: Division of Hematology/Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT

Background: TSC2-deficient cells can proliferate in the lungs, kidneys, and other organs causing devastating progressive multisystem disorders such as lymphangioleiomyomatosis (LAM) and tuberous sclerosis complex (TSC). Preclinical models utilizing LAM patient-derived cells have been difficult to establish. We developed a novel animal model system to study the molecular mechanisms of TSC/LAM pathogenesis and tumorigenesis and provide a platform for drug testing.

Methods and findings: TSC2-deficient human cells, derived from the angiomyolipoma of a LAM patient, were engineered to co-express both sodium-iodide symporter (NIS) and green fluorescent protein (GFP). Cells were inoculated intraparenchymally, intravenously, or intratracheally into athymic NCr nu/nu mice and cells were tracked and quantified using single photon emission computed tomography (SPECT) and computed tomography (CT). Surprisingly, TSC2-deficient cells administered intratracheally resulted in rapid dissemination to lymph node basins throughout the body, and histopathological changes in the lung consistent with LAM. Estrogen was found to be permissive for tumor growth and dissemination. Rapamycin inhibited tumor growth, but tumors regrew after the drug treatment was withdrawn.

Conclusions: We generated homogeneous NIS/GFP co-expressing TSC2-deficient, patient-derived cells that can proliferate and migrate in vivo after intratracheal instillation. Although the animal model we describe has some limitations, we demonstrate that systemic tumors formed from TSC2-deficient cells can be monitored and quantified noninvasively over time using SPECT/CT, thus providing a much needed model system for in vivo drug testing and mechanistic studies of TSC2-deficient cells and their related clinical syndromes.

Show MeSH
Related in: MedlinePlus