Limits...
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 Rapamycin inhibited tumor growth, but tumors regrew after the drug treatment was withdrawn.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.

Show MeSH

Related in: MedlinePlus

Development of a quantifiable LAM/TSC animal model system.A. SPECT/CT imaging of NCr nu/nu mice 4 weeks after inoculation of 621-327 cells by intravenous injection (left), intrapulmonary injection (middle), and intratracheal instillation (right). MIP  =  maximal intensity projection; T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors confirmed after animal sacrifice. Scale bars = 1 cm. *Two of four mice died immediately after direct lung injections. †One mouse showed radiotracer uptake at week 4; others varied from week 8 to week 20. B. SPECT/CT imaging (MIP) of a single mouse preinoculation (Day 0) and at varying times after intratracheal delivery of 621-327 cells. T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors. Scale bars = 1 cm. C. Kinetics of tumor growth measured using SPECT. Shown is %ID/mm3*10−5 (mean ± SD) in 6 tumors per mouse for n = 3 mice over time.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3376142&req=5

pone-0038589-g002: Development of a quantifiable LAM/TSC animal model system.A. SPECT/CT imaging of NCr nu/nu mice 4 weeks after inoculation of 621-327 cells by intravenous injection (left), intrapulmonary injection (middle), and intratracheal instillation (right). MIP  =  maximal intensity projection; T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors confirmed after animal sacrifice. Scale bars = 1 cm. *Two of four mice died immediately after direct lung injections. †One mouse showed radiotracer uptake at week 4; others varied from week 8 to week 20. B. SPECT/CT imaging (MIP) of a single mouse preinoculation (Day 0) and at varying times after intratracheal delivery of 621-327 cells. T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors. Scale bars = 1 cm. C. Kinetics of tumor growth measured using SPECT. Shown is %ID/mm3*10−5 (mean ± SD) in 6 tumors per mouse for n = 3 mice over time.

Mentions: LAM is an unusual disease because the tumor cells have a benign histological phenotype, but also have metastatic potential. To determine whether TSC2-deficient 621-327 cells are capable of disseminating and metastasizing in vivo, we administrated cells into mice by intravenous injection, direct lung injection, or intratracheal instillation. Interestingly, similar tumor cell deposition patterns were seen (Figure 2A), demonstrating that the TSC2-deficient cells in vivo retained tumorigenicity and were disseminated systemically, although the tumor take rate and latency varied in different administrations. Intravenous administration of cells into athymic NCr nu/nu mice resulted in a long latency and low take rate, with tumors also trending toward a smaller size compared with other modes of administration (Figure 2A). Direct needle injection of cells into the lung resulted in distant tumor growth, but also high mortality. Delivery of 621-327 cells via intratracheal instillation resulted in tumor growth in 16 of 16 animals, short latency, widespread dissemination, and relatively large tumors.


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)

Development of a quantifiable LAM/TSC animal model system.A. SPECT/CT imaging of NCr nu/nu mice 4 weeks after inoculation of 621-327 cells by intravenous injection (left), intrapulmonary injection (middle), and intratracheal instillation (right). MIP  =  maximal intensity projection; T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors confirmed after animal sacrifice. Scale bars = 1 cm. *Two of four mice died immediately after direct lung injections. †One mouse showed radiotracer uptake at week 4; others varied from week 8 to week 20. B. SPECT/CT imaging (MIP) of a single mouse preinoculation (Day 0) and at varying times after intratracheal delivery of 621-327 cells. T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors. Scale bars = 1 cm. C. Kinetics of tumor growth measured using SPECT. Shown is %ID/mm3*10−5 (mean ± SD) in 6 tumors per mouse for n = 3 mice over time.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038589-g002: Development of a quantifiable LAM/TSC animal model system.A. SPECT/CT imaging of NCr nu/nu mice 4 weeks after inoculation of 621-327 cells by intravenous injection (left), intrapulmonary injection (middle), and intratracheal instillation (right). MIP  =  maximal intensity projection; T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors confirmed after animal sacrifice. Scale bars = 1 cm. *Two of four mice died immediately after direct lung injections. †One mouse showed radiotracer uptake at week 4; others varied from week 8 to week 20. B. SPECT/CT imaging (MIP) of a single mouse preinoculation (Day 0) and at varying times after intratracheal delivery of 621-327 cells. T  =  thyroid; S  =  stomach; B  =  bladder. Arrows indicate tumors. Scale bars = 1 cm. C. Kinetics of tumor growth measured using SPECT. Shown is %ID/mm3*10−5 (mean ± SD) in 6 tumors per mouse for n = 3 mice over time.
Mentions: LAM is an unusual disease because the tumor cells have a benign histological phenotype, but also have metastatic potential. To determine whether TSC2-deficient 621-327 cells are capable of disseminating and metastasizing in vivo, we administrated cells into mice by intravenous injection, direct lung injection, or intratracheal instillation. Interestingly, similar tumor cell deposition patterns were seen (Figure 2A), demonstrating that the TSC2-deficient cells in vivo retained tumorigenicity and were disseminated systemically, although the tumor take rate and latency varied in different administrations. Intravenous administration of cells into athymic NCr nu/nu mice resulted in a long latency and low take rate, with tumors also trending toward a smaller size compared with other modes of administration (Figure 2A). Direct needle injection of cells into the lung resulted in distant tumor growth, but also high mortality. Delivery of 621-327 cells via intratracheal instillation resulted in tumor growth in 16 of 16 animals, short latency, widespread dissemination, and relatively large tumors.

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