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A standardized autopsy procurement allows for the comprehensive study of DIPG biology.

Kambhampati M, Perez JP, Yadavilli S, Saratsis AM, Hill AD, Ho CY, Panditharatna E, Markel M, Packer RJ, Nazarian J - Oncotarget (2015)

Bottom Line: Diffuse intrinsic pontine glioma (DIPG) is one of the least understood and most deadly childhood cancers.Collaborative efforts to share data and tumor specimens have resulted in rapid discoveries in other pediatric brain tumors, such as medulloblastoma, and therefore have the potential to shed light on the biology of DIPG.Sixteen autopsies were performed throughout the United States and Canada and processed using a standard protocol and inventory method, including specimen imaging, fixation, snap freezing, orthotopic injection, or preservation.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Genetic Medicine, Children's National Health System, Washington, DC, USA.

ABSTRACT
Diffuse intrinsic pontine glioma (DIPG) is one of the least understood and most deadly childhood cancers. Historically, there has been a paucity of DIPG specimens for molecular analysis. However, due to the generous participation of DIPG families in programs for postmortem specimen donation, there has been a recent surge in molecular analysis of newly available tumor specimens. Collaborative efforts to share data and tumor specimens have resulted in rapid discoveries in other pediatric brain tumors, such as medulloblastoma, and therefore have the potential to shed light on the biology of DIPG. Given the generous gift of postmortem tissue donation from DIPG patients, there is a need for standardized postmortem specimen accrual to facilitate rapid and effective multi-institutional molecular studies.We developed and implemented an autopsy protocol for rapid procurement, documenting and storing these specimens. Sixteen autopsies were performed throughout the United States and Canada and processed using a standard protocol and inventory method, including specimen imaging, fixation, snap freezing, orthotopic injection, or preservation. This allowed for comparative clinical and biological studies of rare postmortem DIPG tissue specimens, generation of in vivo and in vitro models of DIPG, and detailed records to facilitate collaborative analysis.

No MeSH data available.


Related in: MedlinePlus

Orthotopic injection of DMSO preserved cells results in diffuse pontine tumors(a) Mice were injected orthotopically in brainstem with cells stored in DMSO. Mice that showed signs of tumor development were sacrificed, brains were fixed and processed for histological studies. H&E staining showed tumor formation in pons (P) as judged by (b) hypercellularity, (c) proliferation marker Ki67, (d) human mitochondrial protein MAB1273 (e) histone 3 K27 trimethylation, and (f) histone 3 K27M mutation.
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Figure 5: Orthotopic injection of DMSO preserved cells results in diffuse pontine tumors(a) Mice were injected orthotopically in brainstem with cells stored in DMSO. Mice that showed signs of tumor development were sacrificed, brains were fixed and processed for histological studies. H&E staining showed tumor formation in pons (P) as judged by (b) hypercellularity, (c) proliferation marker Ki67, (d) human mitochondrial protein MAB1273 (e) histone 3 K27 trimethylation, and (f) histone 3 K27M mutation.

Mentions: Fresh autopsy specimens were stored in DMSO as described in Materials and Methods.. DMSO preserved specimens (10 hour post mortem) from one patient were orthotopically injected into the brainstem of ten-day old (p10) mice (n=10). Injections were performed according to established methods [7] by hand at 2 mm posterior to the bregma at the midline position. Seventy percent of injected mice resulted in infiltrating pontine (Fig. 5a, b) or cortical (data not shown) tumors positive for proliferative marker Ki67 (Fig. 5c ). These tumors were also positive for human mitochondrial protein as detected by MAB1273 antibody (Fig. 5d). Tumors showed reduced Histone 3 K27 trimethylation staining (Fig. 5e) but increased Histone 3 K27 mutation (Fig. 5f) indicating their human cell of origin.


A standardized autopsy procurement allows for the comprehensive study of DIPG biology.

Kambhampati M, Perez JP, Yadavilli S, Saratsis AM, Hill AD, Ho CY, Panditharatna E, Markel M, Packer RJ, Nazarian J - Oncotarget (2015)

Orthotopic injection of DMSO preserved cells results in diffuse pontine tumors(a) Mice were injected orthotopically in brainstem with cells stored in DMSO. Mice that showed signs of tumor development were sacrificed, brains were fixed and processed for histological studies. H&E staining showed tumor formation in pons (P) as judged by (b) hypercellularity, (c) proliferation marker Ki67, (d) human mitochondrial protein MAB1273 (e) histone 3 K27 trimethylation, and (f) histone 3 K27M mutation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494970&req=5

Figure 5: Orthotopic injection of DMSO preserved cells results in diffuse pontine tumors(a) Mice were injected orthotopically in brainstem with cells stored in DMSO. Mice that showed signs of tumor development were sacrificed, brains were fixed and processed for histological studies. H&E staining showed tumor formation in pons (P) as judged by (b) hypercellularity, (c) proliferation marker Ki67, (d) human mitochondrial protein MAB1273 (e) histone 3 K27 trimethylation, and (f) histone 3 K27M mutation.
Mentions: Fresh autopsy specimens were stored in DMSO as described in Materials and Methods.. DMSO preserved specimens (10 hour post mortem) from one patient were orthotopically injected into the brainstem of ten-day old (p10) mice (n=10). Injections were performed according to established methods [7] by hand at 2 mm posterior to the bregma at the midline position. Seventy percent of injected mice resulted in infiltrating pontine (Fig. 5a, b) or cortical (data not shown) tumors positive for proliferative marker Ki67 (Fig. 5c ). These tumors were also positive for human mitochondrial protein as detected by MAB1273 antibody (Fig. 5d). Tumors showed reduced Histone 3 K27 trimethylation staining (Fig. 5e) but increased Histone 3 K27 mutation (Fig. 5f) indicating their human cell of origin.

Bottom Line: Diffuse intrinsic pontine glioma (DIPG) is one of the least understood and most deadly childhood cancers.Collaborative efforts to share data and tumor specimens have resulted in rapid discoveries in other pediatric brain tumors, such as medulloblastoma, and therefore have the potential to shed light on the biology of DIPG.Sixteen autopsies were performed throughout the United States and Canada and processed using a standard protocol and inventory method, including specimen imaging, fixation, snap freezing, orthotopic injection, or preservation.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Genetic Medicine, Children's National Health System, Washington, DC, USA.

ABSTRACT
Diffuse intrinsic pontine glioma (DIPG) is one of the least understood and most deadly childhood cancers. Historically, there has been a paucity of DIPG specimens for molecular analysis. However, due to the generous participation of DIPG families in programs for postmortem specimen donation, there has been a recent surge in molecular analysis of newly available tumor specimens. Collaborative efforts to share data and tumor specimens have resulted in rapid discoveries in other pediatric brain tumors, such as medulloblastoma, and therefore have the potential to shed light on the biology of DIPG. Given the generous gift of postmortem tissue donation from DIPG patients, there is a need for standardized postmortem specimen accrual to facilitate rapid and effective multi-institutional molecular studies.We developed and implemented an autopsy protocol for rapid procurement, documenting and storing these specimens. Sixteen autopsies were performed throughout the United States and Canada and processed using a standard protocol and inventory method, including specimen imaging, fixation, snap freezing, orthotopic injection, or preservation. This allowed for comparative clinical and biological studies of rare postmortem DIPG tissue specimens, generation of in vivo and in vitro models of DIPG, and detailed records to facilitate collaborative analysis.

No MeSH data available.


Related in: MedlinePlus