eNews Core Spotlight:
Specialized Histopathology Core, MGH Site
JULY 27, 2017
In this DF/HCC Core Spotlight, we highlight the Specialized Histopathology Core which has both an MGH and a Longwood site, and in this edition we will focus specifically on the MGH site which is located in the Charlestown Navy Yard. The Core provides technical and professional pathology services to investigators conducting research with experimental organisms (e.g. rodents, fish, and non-human primates) or human tissues, including assistance with experimental design and data interpretation expertise.
We have now added automated Immunohistochemistry (Ventana) service :
- Optimization of novel antibodies
- Dual staining
Services provided at the Core's MGH site include:
- Tissue trimming, cassetting, processing, and embedding;
- Cutting paraffin-embedded and cryostat sections;
- H&E, special stains, TUNEL
- Optimization of new markers
- Immunostaining with one or two markers manually
- Immunostaining with one or two markers with automated immunostainer (Ventana)
Optimizations and slides are often reviewed and discussed with the researchers, ensuring continuity and the development of productive working relationship with the staff. The core director, Anat Stemmer-Rachamimov, MD (MGH) is a neuropathologist, and in addition to the general services listed above provides expertise in neuropathology and eye pathology (human and animal models).
The core is located in Charlestown, Building 149, but pickup and drop off of slides as well as slide review can be arranged in MGH, if requested.
In vivo imaging of tumor-propagating cells, regional tumor heterogeneity and dynamic cell movements in embryonal rhabdomyosarcoma
Principal Investigator: David LangenauMGH (Leukemia)Funding Source: American Cancer Society and the NCI (R24OD016761 and 1R01CA154923)
This study used a transgenic zebra fish model of embryonal rhabdomyosarcoma (ERMS) to analyze tumor heterogeneity and the role of different cell populations in tumor dissemination. Using immunohistochemistry, the study confirmed the existence of molecularly definable ERMS-propagating cell that expresses myf5, m-cadherin, and c-met and lacks markers of more differentiated muscle cells. These myf5+ ERMS-propagating cells exist in niches within the tumor, while more differentiated myogenin+
ERMS cells derived from these cells show migratory capability and move away from the ERMS-propagating cells of origin. This work suggests that metastatic capacity and tumor-propagating potential reside in distinct cell population. The Core supported the work by performing tissue embedding and processing, slide preparation, H&E staining, and optimization and IHC staining of myogenin, PAX7, and MyoD (in zebra fish and human).
Publication: Ignatius MS, Chen E, Elpek NM, Fuller AZ, Tenente IM, Clagg R, Liu S, Blackburn JS, Linardic CM, Rosenberg AE, Nielsen PG, Mempel TR, Langenau DMMGH. In vivo imaging of tumor-propagating cells, regional tumor heterogeneity and dynamic cell movements in embryonal rhabdomyosarcoma. Cancer Cell, 2012, 21(5):680-93.
Cancer therapy. Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility
Principal Investigator: Haber DAMGH (Cancer Cell Biology, Cancer Genetics)
Funding Source: Breast Cancer Research Foundation (Haber DA), Stand Up to Cancer (Haber DA, Toner M, Maheswaran S.), National Foundation for Cancer Research (Haber DA.), NIH CA129933 (Haber DA.), Susan G. Komen for the Cure KG09042 (Maheswaran S.), National Cancer Institute–MGH Proton Federal Share Program (Maheswaran S), the Howard Hughes Medical Institute (Yu M and Haber DA).
This is a study involving multiple members in MGH (Bardia A, Sgroi D, Getz G, Iafrate AJ, Maheswaran S, and Haber DA) in which circulating tumor cells (CTC) from patients with estrogen receptor positive breast cancer have been isolated and established in culture. Three of five CTC lines tested were tumorigenic in mice. Genome sequencing revealed both pre-existing mutations and newly acquired mutations. This is a proof of concept study which provides support to a novel strategy of individualized medicine: utilization of isolated circulating tumor cells to identify novel mutations and to perform drug screenings. This strategy may help identify the best suited therapies for individuals with cancer over the course of their disease. The Core provided embedding, tissue processing, slide preparation, H&E staining, and IHC.
Publication: Yu M, Bardia A, Aceto N, Bersani F, Madden MN, Donaldson MC, Desai R, Comaills V, Zheng Z, Brachtel E, Sgroi D, Kupur R, Getz G, Iafrate AJ, Benes C, Toner M, Maheswaran S, and Haber DA. Cancer therapy. Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility. Science. 2014, 345(6193):216-20 PMID: 25013076
(A) Representative images of nonadherent CTC culture (BRx-07). Top: Phase contrast. Scale bar, 100 μm. Middle: immunofluorescent staining for cytokeratin (CK, red), Ki67 (yellow), CD45 (green), nuclei [4′,6-diamidino-2-phenylindole (DAPI), blue]. Scale bar, 20 μm. Bottom: Light microscopic imaging with Papanicolaou staining. Comparable images for uncultured primary CTCs are shown in the insets. Scale bar, 20 μm.
(B) (Left) Bioluminescent images showing growth of NSG mouse xenografts, after implantation of 20,000 cultured CTCs (BRx-07) into the mammary fat pad. (Right) Quantification of bioluminescent signals for BRx-07–derived mouse xenografts (mean ± SD, n = 6).
(C) Histology of matched primary breast tumors, cultured CTCs, and CTC-derived mouse xenografts for two CTC lines. All panels show cellular staining with hematoxylin (blue) and immunohistochemical staining for ER expression (brown). Scale bar, 20 μm.
For more information or to contact the Specialized Histopathology Core, visit the core website here.