1) Using high resolution telomere fluorescent in situ hybridization, the study found that intratubular germ cell neoplasia and most pure seminomas exhibited short telomeres, while non-seminoma subtypes generally had longer telomeres and a worse prognosis.
2) Among non-seminomas, embryonal carcinoma had the highest proportion of long telomeres and was the only subtype to display an alternative lengthening of telomeres phenotype.
3) The findings indicate that telomere anomalies are an early event in testicular germ cell tumor carcinogenesis and may contribute to tumor initiation and progression. Further investigation of telomeric aberrations could provide new pathways for
Osteoblasts remotely supply lung tumors with cancer-promoting SiglecFhigh neu...Gul Muneer
Systemic cross-talk between lung tumors and bones
Bone marrow–derived myeloid cells can accumulate within tumors and foster
cancer outgrowth. Local immune-neoplastic interactions have been intensively
investigated, but the contribution of the systemic host environment to tumor growth
remains poorly understood. Here, we show in mice and cancer patients (n = 70) that
lung adenocarcinomas increase bone stromal activity in the absence of bone
metastasis. Animal studies reveal that the cancer-induced bone phenotype involves
bone-resident osteocalcin-expressing (Ocn+) osteoblastic cells. These cells promote
cancer by remotely supplying a distinct subset of tumor-infiltrating SiglecFhigh
neutrophils, which exhibit cancer-promoting properties. Experimentally reducing
Ocn+ cell numbers suppresses the neutrophil response and lung tumor outgrowth.
These observations posit osteoblasts as remote regulators of lung cancer and
identify SiglecFhigh neutrophils as myeloid cell effectors of the osteoblast-driven
protumoral response
Proteogenomic analysis of human colon cancer reveals new therapeutic opportun...Gul Muneer
We performed the first proteogenomic study on a prospectively collected colon cancer cohort. Comparative proteomic and phosphoproteomic analysis of paired tumor and normal adjacent tissues produced a catalog of colon cancer-associated proteins and phosphosites, including known and putative new biomarkers, drug targets, and cancer/testis antigens. Proteogenomic integration not only prioritized genomically inferred targets, such as copy-number drivers and mutation-derived neoantigens, but also yielded novel findings. Phosphoproteomics data associated Rb phosphorylation with increased proliferation and decreased apoptosis in colon cancer, which explains why this classical tumor suppressor is amplified in colon tumors and suggests a rationale for targeting Rb phosphorylation in colon cancer. Proteomics identified an association between decreased CD8 T cell infiltration and increased glycolysis in microsatellite instability-high (MSI-H) tumors, suggesting glycolysis as a potential target to overcome the resistance of MSI-H tumors to immune checkpoint blockade. Proteogenomics presents new avenues for biological discoveries and therapeutic development.
Assessing the clinical utility of cancer genomic and proteomic data across tu...Gul Muneer
Molecular profiling of tumors promises to advance the clinical
management of cancer, but the benefits of integrating
molecular data with traditional clinical variables have not been
systematically studied. Here we retrospectively predict patient
survival using diverse molecular data (somatic copy-number
alteration, DNA methylation and mRNA, microRNA and protein
expression) from 953 samples of four cancer types from The
Cancer Genome Atlas project. We find that incorporating
molecular data with clinical variables yields statistically
significantly improved predictions (FDR < 0.05) for three
cancers but those quantitative gains were limited (2.2–23.9%).
Additional analyses revealed little predictive power across
tumor types except for one case. In clinically relevant genes,
we identified 10,281 somatic alterations across 12 cancer types
in 2,928 of 3,277 patients (89.4%), many of which would
not be revealed in single-tumor analyses. Our study provides
a starting point and resources, including an open-access
model evaluation platform, for building reliable prognostic and
therapeutic strategies that incorporate molecular data
The use of genetic engineering technology in animals has been associated with ethical issues, some of which relate to animal welfare. Discuss examples of genetically engineering animals and evaluate the ethical concerns of genetic engineering.
Osteoblasts remotely supply lung tumors with cancer-promoting SiglecFhigh neu...Gul Muneer
Systemic cross-talk between lung tumors and bones
Bone marrow–derived myeloid cells can accumulate within tumors and foster
cancer outgrowth. Local immune-neoplastic interactions have been intensively
investigated, but the contribution of the systemic host environment to tumor growth
remains poorly understood. Here, we show in mice and cancer patients (n = 70) that
lung adenocarcinomas increase bone stromal activity in the absence of bone
metastasis. Animal studies reveal that the cancer-induced bone phenotype involves
bone-resident osteocalcin-expressing (Ocn+) osteoblastic cells. These cells promote
cancer by remotely supplying a distinct subset of tumor-infiltrating SiglecFhigh
neutrophils, which exhibit cancer-promoting properties. Experimentally reducing
Ocn+ cell numbers suppresses the neutrophil response and lung tumor outgrowth.
These observations posit osteoblasts as remote regulators of lung cancer and
identify SiglecFhigh neutrophils as myeloid cell effectors of the osteoblast-driven
protumoral response
Proteogenomic analysis of human colon cancer reveals new therapeutic opportun...Gul Muneer
We performed the first proteogenomic study on a prospectively collected colon cancer cohort. Comparative proteomic and phosphoproteomic analysis of paired tumor and normal adjacent tissues produced a catalog of colon cancer-associated proteins and phosphosites, including known and putative new biomarkers, drug targets, and cancer/testis antigens. Proteogenomic integration not only prioritized genomically inferred targets, such as copy-number drivers and mutation-derived neoantigens, but also yielded novel findings. Phosphoproteomics data associated Rb phosphorylation with increased proliferation and decreased apoptosis in colon cancer, which explains why this classical tumor suppressor is amplified in colon tumors and suggests a rationale for targeting Rb phosphorylation in colon cancer. Proteomics identified an association between decreased CD8 T cell infiltration and increased glycolysis in microsatellite instability-high (MSI-H) tumors, suggesting glycolysis as a potential target to overcome the resistance of MSI-H tumors to immune checkpoint blockade. Proteogenomics presents new avenues for biological discoveries and therapeutic development.
Assessing the clinical utility of cancer genomic and proteomic data across tu...Gul Muneer
Molecular profiling of tumors promises to advance the clinical
management of cancer, but the benefits of integrating
molecular data with traditional clinical variables have not been
systematically studied. Here we retrospectively predict patient
survival using diverse molecular data (somatic copy-number
alteration, DNA methylation and mRNA, microRNA and protein
expression) from 953 samples of four cancer types from The
Cancer Genome Atlas project. We find that incorporating
molecular data with clinical variables yields statistically
significantly improved predictions (FDR < 0.05) for three
cancers but those quantitative gains were limited (2.2–23.9%).
Additional analyses revealed little predictive power across
tumor types except for one case. In clinically relevant genes,
we identified 10,281 somatic alterations across 12 cancer types
in 2,928 of 3,277 patients (89.4%), many of which would
not be revealed in single-tumor analyses. Our study provides
a starting point and resources, including an open-access
model evaluation platform, for building reliable prognostic and
therapeutic strategies that incorporate molecular data
The use of genetic engineering technology in animals has been associated with ethical issues, some of which relate to animal welfare. Discuss examples of genetically engineering animals and evaluate the ethical concerns of genetic engineering.
Telomerase Inhibition as Novel Cancer Therapeutic MethodVincensanicko
Telomere in cancerous cells is conserved even after several rounds of cell division. By identifying the responsible protein in this process, i.e. telomerase, researchers have utilised it as a novel target for cancer treatment. So, how is the telomerase targetted? Are you ready to discover the truth?
Nanodroplet processing platform for deep and quantitative proteome profiling ...Gul Muneer
Nanoscale or single-cell technologies are critical for biomedical applications. However, current mass spectrometry (MS)-based proteomic approaches require samples comprising a minimum of thousands of cells to provide in-depth profiling. Here, we report the development of a nanoPOTS (nanodroplet processing in one pot for trace samples) platform for small cell population proteomics analysis. NanoPOTS enhances the efficiency and recovery of sample processing by downscaling processing volumes to <200 nL to minimize surface losses. When combined with ultrasensitive liquid chromatography-MS, nanoPOTS allows identification of ~1500 to ~3000 proteins from ~10 to ~140 cells, respectively. By incorporating the Match Between Runs algorithm of MaxQuant, >3000 proteins are consistently identified from as few as 10 cells. Furthermore, we demonstrate quantification of ~2400 proteins from single human pancreatic islet thin sections from type 1 diabetic and control donors, illustrating the application of nanoPOTS for spatially resolved proteome measurements from clinical tissues.
Exploring chemo-resistance in NSCLC - Dr Martin BarrHannahMcCarthy31
Dr Martin Barr is a Clinical Scientist at St James's Hospital and Adjunct Assistant Professor at Trinity College Dublin. Dr Barr's research interests are chemotherapy resistance in Non-Small Cell Lung Cancer (NSCLC), in vivo and in vitro models, Liquid Biopsy and EGFR-mutant NSCLC.
Dr Una L Fairbrother
Telomere length: a 21st century biomarker" discusses DNA structure and the nature of telomeres. This talk explains the importance of telomere length and the impact of this feature on human health. The talk finishes describing the exciting work being carried out in London Metropolitan University to help develop this measure as a 21st century biomarker.
Direct Sanger CE Sequencing of Individual Ampliseq Cancer Panel Targets from ...Thermo Fisher Scientific
The introduction of defined Ion AmpliSeq™ panels for detection and characterization of actionable mutations occurring in tumor tissue has the potential to revolutionize translational oncology research. The Ion Ampliseq™ cancer hot spot panel version 2 (CHP v2) by Ion Torrent includes 207 actionable sequences from a single target and mutation targets present in 50 genes and the more comprehensive Ion Oncomine™ cancer panel (OCP) developed by Life Technologies Compendia Bioscience™ contains over 2000 mutations. A hallmark of these Ion Torrent Ampliseq cancer panels is the low amount of input DNA needed which is critical when the clinical specimen material is limited such as with fine needle biopsy or FFPE samples. Typically, 10 ng of DNA obtained from these sources is sufficient to produce informative sequencing data. Often, cancer-causing or promoting mutations are detected at relatively low allele frequencies like 10-20 % compared to the major normal allele. Many researchers wish to verify these findings of low frequency mutations by an orthologous method such as traditional dye-fluorescent Sanger sequencing on a capillary electrophoresis (CE) instrument such as the Applied Biosystems 3500 genetic analyzer. To that end, we have developed a workflow that enables the amplification and traditional Sanger sequencing of individual Ion AmpliSeq targets directly from the AmpliSeq library starting material.
The method requires a retainer of 1 μl (~ 5%) of the original AmpliSeq preamplification material. A dilution of this aliquot is used as template source for individualized PCR/sequencing reactions. We show that a random selection of 48 targets from the CHPv2 panel could be successfully amplified and Sanger-sequenced from an Ion Torrent Ampliseq library originally prepared from 10 ng of FFPE
DNA. Furthermore, we show the successful Sanger-re-sequencing of all individual 24 targets covering the TP53 exons from the same sample processed and pre-amplified with the OncoMine AmpliSeq panel.
Taken together, this method will enable researchers to reflex-test potential mutations of interest from very material-limited specimen using Sanger CE sequencing
Analytical performance of a novel next generation sequencing assay for Myeloi...Thermo Fisher Scientific
To support clinical and translational research into precision oncology strategies for myeloid cancers, a next-generation sequencing (NGS) assay was developed to detect common and relevant somatic alterations. To define gene targets that were recurrently altered in myeloid cancers and relevant for clinical and translational research, an extensive survey of investigators at hematology oncology research labs was performed.
Telomerase Inhibition as Novel Cancer Therapeutic MethodVincensanicko
Telomere in cancerous cells is conserved even after several rounds of cell division. By identifying the responsible protein in this process, i.e. telomerase, researchers have utilised it as a novel target for cancer treatment. So, how is the telomerase targetted? Are you ready to discover the truth?
Nanodroplet processing platform for deep and quantitative proteome profiling ...Gul Muneer
Nanoscale or single-cell technologies are critical for biomedical applications. However, current mass spectrometry (MS)-based proteomic approaches require samples comprising a minimum of thousands of cells to provide in-depth profiling. Here, we report the development of a nanoPOTS (nanodroplet processing in one pot for trace samples) platform for small cell population proteomics analysis. NanoPOTS enhances the efficiency and recovery of sample processing by downscaling processing volumes to <200 nL to minimize surface losses. When combined with ultrasensitive liquid chromatography-MS, nanoPOTS allows identification of ~1500 to ~3000 proteins from ~10 to ~140 cells, respectively. By incorporating the Match Between Runs algorithm of MaxQuant, >3000 proteins are consistently identified from as few as 10 cells. Furthermore, we demonstrate quantification of ~2400 proteins from single human pancreatic islet thin sections from type 1 diabetic and control donors, illustrating the application of nanoPOTS for spatially resolved proteome measurements from clinical tissues.
Exploring chemo-resistance in NSCLC - Dr Martin BarrHannahMcCarthy31
Dr Martin Barr is a Clinical Scientist at St James's Hospital and Adjunct Assistant Professor at Trinity College Dublin. Dr Barr's research interests are chemotherapy resistance in Non-Small Cell Lung Cancer (NSCLC), in vivo and in vitro models, Liquid Biopsy and EGFR-mutant NSCLC.
Dr Una L Fairbrother
Telomere length: a 21st century biomarker" discusses DNA structure and the nature of telomeres. This talk explains the importance of telomere length and the impact of this feature on human health. The talk finishes describing the exciting work being carried out in London Metropolitan University to help develop this measure as a 21st century biomarker.
Direct Sanger CE Sequencing of Individual Ampliseq Cancer Panel Targets from ...Thermo Fisher Scientific
The introduction of defined Ion AmpliSeq™ panels for detection and characterization of actionable mutations occurring in tumor tissue has the potential to revolutionize translational oncology research. The Ion Ampliseq™ cancer hot spot panel version 2 (CHP v2) by Ion Torrent includes 207 actionable sequences from a single target and mutation targets present in 50 genes and the more comprehensive Ion Oncomine™ cancer panel (OCP) developed by Life Technologies Compendia Bioscience™ contains over 2000 mutations. A hallmark of these Ion Torrent Ampliseq cancer panels is the low amount of input DNA needed which is critical when the clinical specimen material is limited such as with fine needle biopsy or FFPE samples. Typically, 10 ng of DNA obtained from these sources is sufficient to produce informative sequencing data. Often, cancer-causing or promoting mutations are detected at relatively low allele frequencies like 10-20 % compared to the major normal allele. Many researchers wish to verify these findings of low frequency mutations by an orthologous method such as traditional dye-fluorescent Sanger sequencing on a capillary electrophoresis (CE) instrument such as the Applied Biosystems 3500 genetic analyzer. To that end, we have developed a workflow that enables the amplification and traditional Sanger sequencing of individual Ion AmpliSeq targets directly from the AmpliSeq library starting material.
The method requires a retainer of 1 μl (~ 5%) of the original AmpliSeq preamplification material. A dilution of this aliquot is used as template source for individualized PCR/sequencing reactions. We show that a random selection of 48 targets from the CHPv2 panel could be successfully amplified and Sanger-sequenced from an Ion Torrent Ampliseq library originally prepared from 10 ng of FFPE
DNA. Furthermore, we show the successful Sanger-re-sequencing of all individual 24 targets covering the TP53 exons from the same sample processed and pre-amplified with the OncoMine AmpliSeq panel.
Taken together, this method will enable researchers to reflex-test potential mutations of interest from very material-limited specimen using Sanger CE sequencing
Analytical performance of a novel next generation sequencing assay for Myeloi...Thermo Fisher Scientific
To support clinical and translational research into precision oncology strategies for myeloid cancers, a next-generation sequencing (NGS) assay was developed to detect common and relevant somatic alterations. To define gene targets that were recurrently altered in myeloid cancers and relevant for clinical and translational research, an extensive survey of investigators at hematology oncology research labs was performed.
Critical Role of PET-Scan in Unravelling the Dual Pathology- Review of Litera...AnonIshanvi
Simultaneous presentation of two lymphatic haematological malignancies is extremely rare. Adequate and optimal diagnostic steps including various imaging techniques and histopathological biopsies are required unpin the exact diagnoses to be able to deliver the best management strategies
Critical Role of PET-Scan in Unravelling the Dual Pathology- Review of Litera...daranisaha
Simultaneous presentation of two lymphatic haematological malignancies is extremely rare. Adequate and optimal diagnostic steps including various imaging techniques and histopathological biopsies are required unpin the exact diagnoses to be able to deliver the best management strategies...
Critical Role of PET-Scan in Unravelling the Dual Pathology- Review of Litera...JohnJulie1
Simultaneous presentation of two lymphatic haematological malignancies is extremely rare. Adequate and optimal diagnostic steps including various imaging techniques and histopathological biopsies are required unpin the exact diagnoses to be able to deliver the best management strategies...
Critical Role of PET-Scan in Unravelling the Dual Pathology- Review of Litera...NainaAnon
Simultaneous presentation of two lymphatic haematological malignancies is extremely rare. Adequate and optimal diagnostic steps including various imaging techniques and histopathological biopsies are required unpin the exact diagnoses to be able to deliver the best management strategies...
The simultaneous occurrence of two lymphatic malignancies in
one patient is extremely rare with an incidence rate of 1.4–6.5
cases/1,000,000 individuals [8]. Co-existence of MM and other
lymphoid malignancies like Chronic Lymphocytic Leukemia (CLL)
[9], MM and Hodgkin’s Disease (HD) [10], MM and Lympho
Plasmacytic Lymphoma (LPL) [11] has been reported. However,
there are less than 5 reported cases in PubMed of simultaneous
presentation of DLBCL and MM
Critical Role of PET-Scan in Unravelling the Dual Pathology- Review of Litera...EditorSara
Simultaneous presentation of two lymphatic haematological malignancies is extremely rare. Adequate and optimal diagnostic steps including various imaging techniques and histopathological biopsies are required unpin the exact diagnoses to be able to deliver the best management strategies...
Critical Role of PET-Scan in Unravelling the Dual Pathology- Review of Litera...semualkaira
Simultaneous presentation of two lymphatic haematological malignancies is extremely rare. Adequate and optimal diagnostic steps including various imaging techniques and histopathological biopsies are required unpin the exact diagnoses to be able to deliver the best management strategies...
Critical Role of PET-Scan in Unravelling the Dual Pathology- Review of Litera...semualkaira
Simultaneous presentation of two lymphatic haematological malignancies is extremely rare. Adequate and optimal diagnostic steps including various imaging techniques and histopathological biopsies are required unpin the exact diagnoses to be able to deliver the best management strategies...
A normal cell can be transformed into a cancerous cell. Discuss the therapeutic strategies that are employed to target the cellular transformation process for cancer prevention and treatment.
El 27 de noviembre de 2014, la Fundación Ramón Areces celebró una nueva conferencia del ciclo que organiza sobre 'Envejecimiento, Sociedad y Salud: envejecimiento y enfermedad' en colaboración con el Centro de Estudios del Envejecimiento. En esta ocasión, la investigadora María Blasco, directora del Centro Nacional de Investigaciones Oncológicas (CNIO), habló sobre 'El origen de la enfermedad'.
Assessment of immunomolecular_expression_and_prognostic_role_of_tlr7_among_pa...dr.Ihsan alsaimary
Dr. ihsan edan abdulkareem alsaimary
PROFESSOR IN MEDICAL MICROBIOLOGY AND MOLECULAR IMMUNOLOGY
ihsanalsaimary@gmail.com
mobile : 009647801410838
university of basrah - college of medicine - basrah -IRAQ
Presentation by Scott Woodman, MD, PhD. Presented at the 2018 Eyes on a Cure: Patient & Caregiver Symposium, hosted by the Melanoma Research Foundation's CURE OM initiative.
Similar to AACR 2015 Poster 04-16-2015 2005 hrs (20)
1. High resolution telomere FISH reveals anomalies in male germ cell
tumor subtypes and precursor lesion
Mohammed Talha Shekhani1, John R. Barber4, Stephania M. Bezerra1, Christopher M. Heaphy1,3, Nilda Gonzalez Roibon1, Leonardo O. Reis1, Gunes Guner1, Corinne E.
Joshu3,4, George J. Netto1,2,3, and Alan K. Meeker1,2,3
1Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD; 2Department of Urology, James Buchanan Brady Urological Institute at Johns Hopkins, Baltimore, MD; 3Department of
Oncology, Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD; 4Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD
Methodolgy
Testicular germ cell tumor (TGCT) is the most common malignancy of young men between 15-40 years of age. It is biologically
unique, because it is in cells of non-somatic lineage and is an unusually curable cancer.
We evaluated over 70 specimens (arranged on two TGCT tissue microarrays (TMAs)) using high resolution (single-cell) telomere-
specific fluorescent in situ hybridization (FISH) to delineate telomere length patterns in different testicular germ cell tumor
subtypes, as well as in their precursor lesion, intratubular germ cell neoplasia unclassified type (IGCNU). Telomere biology has
never previously been addressed in IGCNU. Oct4 is a highly specific histological marker of IGCNU, seminoma, and embryonal
carcinoma neoplastic cells. We used co-immunofluorescence against Oct4 to improve identification of cancer cells in said
lesions.
Telomere lengths were scored as short (=1), medium (=2) or long (=3) by comparing the intensity of the telomere signals from
neoplastic cells to benign reference cell types. Reference cells are interstitial/stromal cells (short telomeres) and normal germ
cells (medium telomeres).
Abstract
Number:
3783
Summary of Key Findings
Compared with normal germ cells, short telomeres were observed in intratubular germ cell neoplasia (IGCNU) lesions and in majority of pure seminomas
(p=0.006 and p=0.0005, respectively). In contrast, we noted longer telomere lengths in non-seminomas—cancer subtypes that also carry a worse prognosis.
Non-seminoma and seminoma components in mixed subtype demonstrated telomeres with slight variation from pure counterparts. Among the non-seminomas,
EC has the highest proportion of cases scored as having long telomeres.
EC is the only subtype to display telomerase-independent alternative lengthening of telomeres (ALT) phenotype (43% of pure embryonal carcinomas). Loss of
ATRX or DAXX expression is strongly associated with ALT. However, neither protein anomalies existed in sub-group of EC evaluated by IHC—suggesting that there
may be (as yet undiscovered) genetic events associated with ALT in embryonal carcinoma.
The pattern of telomere lengths among TGCT and their precursor, IGCNU, indicate that telomeres are affected early in carcinogenesis. Moreover, telomere
anomalies may contribute to tumor initiation and progression.
Further investigation of telomeric aberration in TGCT may delineate new pathways for intervention and prognostication in this unique malignancy.
Table 2. Telomere length distributions for TGCT subtypes and IGCNU
precursor
Embryonal
Carcinoma
Seminoma Teratoma
Yolk Sac
Tumor
Mixed
Cancer cell
telomere lengths
Long (%) 42.9 14 0 33.3 N/A
Medium (%) 14.3 27.9 100 0 N/A
Short (%) 0 58.1 0 66.7 N/A
ALT Present (%) 42.9 0 0 0 15
IGCNU Present
(%)
71.4 58.1 0 33.3 70
IGCNU telomere
length
Long (%) 0 8 0 0 0
Medium (%) 0 8 0 0 7.1
Short (%) 100 84 0 100 92.9
ALT = alternative lengthening of telomeres; and IGCNU = intratubular
germ cell neoplasia of unclassified type
Table 3. Telomere lengths in mixed germ cell tumors
Embryonal
Carcinoma
Seminoma Teratoma
Yolk Sac
Tumor
N 18 9 12 9
Long (%) 38.9 66.7 0 0
Medium (%) 38.9 11.1 25 77.8
Short (%) 5.6 22.2 75 22.2
ALT (%) 16.7 0 0 0
ALT = alternative lengthening of telomeres
Figure 1. Example image a healthy seminiferous tubule and an adjacent
IGCNU/diseased tubule. (A) DAPI/Nuclear Stain/Blue and
Cy3/Telomere-FISH/Red channels. Telomeres are punctate red dots
within nuclei. Filled/solid arrow indicates neoplastic germ cell with
short telomeres (nearly absent FISH signal) in the IGCNU tubule. Open-
headed arrow indicates healthy spermatogonial stem cell’s nucleus for
comparison. (B) 3 channel (DAPI/Nuclear Stain/Blue, Cy3/Telomere-
FISH/Red and Oct4 immunostaining/Green) merged image. *(Asterisk)
= Region displaying non-specific autofluorescence (often associated
with red blood cell fragments from nearby capillaries); IGCNU =
Intratubular Germ Cell Neoplasia of Unclassified type; FISH =
fluorescence in situ hybridization; and DAPI = 4',6-diamidino-2-
phenylindole. Original magnification is 200X, scale bar is 100
micrometers.
Figure 2. Seminoma cells compared to normal healthy germ
cells from the same patient. (A) and (B) Display DAPI/Nuclear
Stain/Blue and Cy3/Telomere-FISH/Red channels. Telomeres
are punctate red dots within nuclei. (A) Shows region
containing seminoma cells and (B) shows contents of a
healthy unaffected seminiferous tubule. Insets demonstrate
enlarged cells for unequivocal telomere length comparisons.
(C) and (D) Display 3 channel (DAPI/Nuclear Stain/Blue,
Cy3/Telomere-FISH/Red and Oct4 immunostaining /Green)
merged images of (A) and (B), respectively. *(Asterisk) =
Region displaying non-specific autofluorescence; FISH =
fluorescence in situ hybridization; and DAPI = 4',6-diamidino-
2-phenylindole. Original magnification is 400X, scale bar is 50
micrometers.
Figure 3. Comparison of an embryonal carcinoma (A, top) with another
EC displaying ALT phenotype (B, below). Both panels demonstrate 3
channel (DAPI/Nuclear Stain/Blue, Cy3/Telomere-FISH/Red and Oct4
immunostaining/Green) merged image. (A) This EC shows long
telomeres in the cancer cells compared to the adjacent stroma. Inset
delineates dramatic telomere length difference between EC and a
stromal cell. (B) This ALT positive EC shows ultra-bright telomeric DNA
foci (arrows) and intracellular telomere length heterogeneity that is
characteristic. Note short telomeres in surrounding interstitium.
*(Asterisk) = Region displaying non-specific autofluorescence; ALT =
alternative lengthening of telomeres; EC = embryonal carcinoma; FISH =
fluorescence in situ hybridization; and DAPI = 4',6-diamidino-2-
phenylindole. Original magnification is 400X, scale bar is 50
micrometers.