Generalized approach for accelerated maximum likelihood based image restoration applied to three‐dimensional fluorescence microscopy

Schaefer, Lutz H.; Schuster, Dietwald; Herz, H.

doi:10.1046/j.1365-2818.2001.00949.x

Cited by 99 publications

(83 citation statements)

References 20 publications

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“…Axiovision 4.5 software (Carl Zeiss, Toronto, ON, Canada) and a constrained iterative algorithm were used for deconvolution. 29 ChromoAnalyze software (see, quantitative analysis of chromosomal overlaps) was used to determine the presence of translocations in the interphases cells. The metaphases were scored visually for abnormalities.…”

Section: Fluorescent In Situ Hybridization Of Metaphases and Interphasesmentioning

confidence: 99%

Dynamic chromosomal rearrangements in Hodgkin's lymphoma are due to ongoing three-dimensional nuclear remodeling and breakage-bridge-fusion cycles

Guffei¹,

Sarkar²,

Klewes³

et al. 2010

Haematologica

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The online version of this article has a Supplementary Appendix. BackgroundHodgkin's lymphoma is characterized by the presence of mono-nucleated Hodgkin cells and bi-to multi-nucleated Reed-Sternberg cells. We have recently shown telomere dysfunction and aberrant synchronous/asynchronous cell divisions during the transition of Hodgkin cells to Reed-Sternberg cells. 1 Design and MethodsTo determine whether overall changes in nuclear architecture affect genomic instability during the transition of Hodgkin cells to Reed-Sternberg cells, we investigated the nuclear organization of chromosomes in these cells. ResultsThree-dimensional fluorescent in situ hybridization revealed irregular nuclear positioning of individual chromosomes in Hodgkin cells and, more so, in Reed-Sternberg cells. We characterized an increasingly unequal distribution of chromosomes as mono-nucleated cells became multi-nucleated cells, some of which also contained chromosome-poor 'ghost' cell nuclei. Measurements of nuclear chromosome positions suggested chromosome overlaps in both types of cells. Spectral karyotyping then revealed both aneuploidy and complex chromosomal rearrangements: multiple breakage-bridge-fusion cycles were at the origin of the multiple rearranged chromosomes. This conclusion was challenged by super resolution three-dimensional structured illumination imaging of Hodgkin and Reed-Sternberg nuclei. Three-dimensional super resolution microscopy data documented inter-nuclear DNA bridges in multi-nucleated cells but not in mono-nucleated cells. These bridges consisted of chromatids and chromosomes shared by two Reed-Sternberg nuclei. The complexity of chromosomal rearrangements increased as Hodgkin cells developed into multi-nucleated cells, thus indicating tumor progression and evolution in Hodgkin's lymphoma, with Reed-Sternberg cells representing the highest complexity in chromosomal rearrangements in this disease. ConclusionsThis is the first study to demonstrate nuclear remodeling and associated genomic instability leading to the generation of Reed-Sternberg cells of Hodgkin's lymphoma. We defined nuclear remodeling as a key feature of Hodgkin's lymphoma, highlighting the relevance of nuclear architecture in cancer.Key words: 3D nucleus, chromosomal rearrangements, SKY, 3D-SIM, BBF-cycles, ReedSternberg cells.Citation: Guffei A, Sarkar R, Klewes L, Righolt C, Knecht H, and Mai S. Dynamic chromosomal rearrangements in Hodgkin's lymphoma are due to ongoing three-dimensional nuclear remodeling and breakage-bridge-fusion cycles. Haematologica 2010;95(12):2038-2046. doi:10.3324/haematol.2010 Dynamic chromosomal rearrangements in Hodgkin's lymphoma are due to ongoing three-dimensional nuclear remodeling and breakage-bridge-fusion cycles © F e r r a t a S t o r t i F o u n d a t i o n

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Section: Fluorescent In Situ Hybridization Of Metaphases and Interphasesmentioning

confidence: 99%

Dynamic chromosomal rearrangements in Hodgkin's lymphoma are due to ongoing three-dimensional nuclear remodeling and breakage-bridge-fusion cycles

Guffei¹,

Sarkar²,

Klewes³

et al. 2010

Haematologica

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“…The most reliable one uses constrained iterative deconvolution [Schaefer et al, 2001;Vermolen et al, 2005b]. Telomere measurements are done with a special algorithm and software package that we developed, TeloView TM [Chuang et al, 2004;Vermolen et al, 2005a].…”

Section: Measurement Of Telomere Dysfunction (3d Volumes and Positions)mentioning

confidence: 99%

The significance of telomeric aggregates in the interphase nuclei of tumor cells

Mai

Garini

2006

J of Cellular Biochemistry

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“…Images were deconvolved using a constrained iterative algorithm (Schaefer et al, 2001). Analysis was performed with TeloView (Chuang et al, 2004;Vermolen et al, 2005).…”

Section: Facs Analysismentioning

confidence: 99%

Telomeric aggregates and end-to-end chromosomal fusions require myc box II

et al. 2006

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Telomeres of tumor cells form telomeric aggregates (TAs) within the three-dimensional (3D) interphase nucleus. Some of these TAs represent end-to-end chromosomal fusions and may subsequently initiate breakage-bridgefusion cycles. Wild-type (wt) and myc box II mutant (mt) Myc induce different types of genomic instability when conditionally expressed in mouse proB cells (Ba/F3). Only wt Myc overexpressing Ba/F3 cells are capable of tumor formation in severe combined immunodeficient mice. In this study, we investigated whether telomere dysfunction leading to TA formation is linked to the genetic changes that permit wt c-Myc-dependent transformation of Ba/F3 cells. To this end, we examined the 3D organization of telomeres after the deregulated expression of deletion myc boxII mutant (D106) or wt Myc. D106-Myc overexpression did not induce TAs, whereas wt-Myc deregulation did. Instead, D106-Myc remodelled the 3D telomeric organization such that telomeres aligned in the center of the 3D interphase nucleus forming a telomeric disk owing to a D106-induced G1/S cell cycle arrest. In contrast, wtMyc overexpression led to distorted telomere distribution and TA formation. Analysis of chromosomal alterations using spectral karyotyping confirmed D106-Myc and wtMyc-associated genomic instability. A significant number of chromosomal end-to-end fusions indicative of telomere dysfunction were noted in wt-Myc-expressing cells only. This study suggests that TAs may play a fundamental role in Myc-induced tumorigenesis and provides a novel way to dissect tumor initiation.

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Generalized approach for accelerated maximum likelihood based image restoration applied to three‐dimensional fluorescence microscopy

Cited by 99 publications

References 20 publications

Dynamic chromosomal rearrangements in Hodgkin's lymphoma are due to ongoing three-dimensional nuclear remodeling and breakage-bridge-fusion cycles

Dynamic chromosomal rearrangements in Hodgkin's lymphoma are due to ongoing three-dimensional nuclear remodeling and breakage-bridge-fusion cycles

The significance of telomeric aggregates in the interphase nuclei of tumor cells

Telomeric aggregates and end-to-end chromosomal fusions require myc box II

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