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Nature Aug 2023Recent studies have documented frequent evolution of clones carrying common cancer mutations in apparently normal tissues, which are implicated in cancer development....
Recent studies have documented frequent evolution of clones carrying common cancer mutations in apparently normal tissues, which are implicated in cancer development. However, our knowledge is still missing with regard to what additional driver events take place in what order, before one or more of these clones in normal tissues ultimately evolve to cancer. Here, using phylogenetic analyses of multiple microdissected samples from both cancer and non-cancer lesions, we show unique evolutionary histories of breast cancers harbouring der(1;16), a common driver alteration found in roughly 20% of breast cancers. The approximate timing of early evolutionary events was estimated from the mutation rate measured in normal epithelial cells. In der(1;16)(+) cancers, the derivative chromosome was acquired from early puberty to late adolescence, followed by the emergence of a common ancestor by the patient's early 30s, from which both cancer and non-cancer clones evolved. Replacing the pre-existing mammary epithelium in the following years, these clones occupied a large area within the premenopausal breast tissues by the time of cancer diagnosis. Evolution of multiple independent cancer founders from the non-cancer ancestors was common, contributing to intratumour heterogeneity. The number of driver events did not correlate with histology, suggesting the role of local microenvironments and/or epigenetic driver events. A similar evolutionary pattern was also observed in another case evolving from an AKT1-mutated founder. Taken together, our findings provide new insight into how breast cancer evolves.
Topics: Adolescent; Adult; Female; Humans; Young Adult; Breast Neoplasms; Cell Lineage; Clone Cells; Epigenesis, Genetic; Epithelial Cells; Epithelium; Evolution, Molecular; Microdissection; Mutagenesis; Mutation; Mutation Rate; Premenopause; Tumor Microenvironment
PubMed: 37495687
DOI: 10.1038/s41586-023-06333-9 -
The American Journal of Forensic... Dec 2019Semen is crucial evidence for some sex crimes, with its sole confirmation being sperm detection. The success of sperm detection is dependent on all levels of preanalytic... (Review)
Review
Semen is crucial evidence for some sex crimes, with its sole confirmation being sperm detection. The success of sperm detection is dependent on all levels of preanalytic and analytic procedures. Specimen collection must be performed by well-trained and competent forensic physicians as well as forensic nurses, with preservation done properly before laboratory transfer. Laboratory procedures should consider archival sperm identification, by visualization, with adequate amounts separated from other cells to obtain male DNA profiles. Differential extraction is robust and accepted as the forensic standard but is time consuming and may result in male DNA loss. Thus, alternative methods and microdevices have been developed. Challenges in sperm isolation from vaginal or buccal epithelium mixes and discrimination in multiperpetrator cases have been overcome by single-cell profiling; however, problems inherent in identical twin discrimination and azoospermia have yet to be solved. Epigenetics and future molecular biomarkers may hold the key; therefore, all laboratory processes must consider DNA and RNA protection. Long-term specimen preservation should be done when possible in light of future confirmatory tests.
Topics: Cell Separation; DNA Fingerprinting; DNA Methylation; Female; Forensic Sciences; Humans; Laser Capture Microdissection; Male; Microsatellite Repeats; Polymerase Chain Reaction; Prostate-Specific Antigen; RNA, Messenger; Seminal Vesicle Secretory Proteins; Sex Offenses; Specimen Handling; Spermatozoa; Staining and Labeling; Time Factors
PubMed: 31687979
DOI: 10.1097/PAF.0000000000000517 -
Proteomics Sep 2020The problem with cancer tissue is that its intratumoral heterogeneity and its complexity is extremely high as cells possess, depending on their location and function,... (Review)
Review
The problem with cancer tissue is that its intratumoral heterogeneity and its complexity is extremely high as cells possess, depending on their location and function, different mutations, different mRNA expression and the highest intricacy in the protein pattern. Prior to genomic and proteomic analyses, it is therefore indispensable to identify the exact part of the tissue or even the exact cell. Laser-based microdissection is a tried and tested technique able to produce pure and well-defined cell material for further analysis with proteomic and genomic techniques. It sheds light on the heterogeneity of cancer or other complex diseases and enables the identification of biomarkers. This review aims to raise awareness for the reconsideration of laser-based microdissection and seeks to present current state-of-the-art combinations with omic techniques.
Topics: Genome; Genomics; Humans; Laser Capture Microdissection; Neoplasms; Proteomics
PubMed: 32578340
DOI: 10.1002/pmic.202000077 -
Current Opinion in Urology Jan 2023To review noteworthy research from the last 2 years on surgical management of azoospermia. (Review)
Review
PURPOSE OF REVIEW
To review noteworthy research from the last 2 years on surgical management of azoospermia.
RECENT FINDINGS
The recommended treatments for nonobstructive and obstructive azoospermia have not appreciably changed. However, recent level-1 evidence has reinforced superiority of micro-dissection testicular sperm extraction over sperm aspiration in men with nonobstructive azoospermia, and several studies have identified genetic and other clinical factors that may aid in selecting candidates for testicular sperm extraction. Machine learning technology has shown promise as a decision support system for patient selection prior to sperm retrieval as well a tool to aid in sperm identification from testis tissue.
SUMMARY
Most men with obstructive azoospermia who desire fertility can be offered either surgical reconstruction or sperm retrieval. For men with nonobstructive azoospermia, sperm retrieval with microdissection testicular sperm extraction remains the gold standard treatment. Uncovering more genetic causes of nonobstructive azoospermia may aid in properly counseling and selecting patients for microdissection testicular sperm extraction. Neural networks and deep learning may have a future role in patient selection for surgical sperm retrieval and postprocedural sperm identification.
Topics: Humans; Male; Azoospermia; Semen; Sperm Retrieval; Microdissection; Testis; Retrospective Studies
PubMed: 36301052
DOI: 10.1097/MOU.0000000000001055 -
International Journal of Urology :... Jan 2024Approximately 1% of the general male population has azoospermia, and nonobstructive azoospermia accounts for the majority of cases. The causes vary widely, including... (Review)
Review
Approximately 1% of the general male population has azoospermia, and nonobstructive azoospermia accounts for the majority of cases. The causes vary widely, including chromosomal and genetic abnormalities, varicocele, drug-induced causes, and gonadotropin deficiency; however, the cause is often unknown. In azoospermia caused by hypogonadotropic hypogonadism, gonadotropin replacement therapy can be expected to produce sperm in the ejaculate. In some cases, upfront varicocelectomy for nonobstructive azoospermia with varicocele may result in the appearance of ejaculated spermatozoa; however, the appropriate indication should be selected. Each guideline recommends microdissection testicular sperm extraction for nonobstructive azoospermia in terms of successful sperm retrieval and avoidance of complications. Sperm retrieval rates generally ranged from 20% to 70% but vary depending on the causative disease. Various attempts have been made to predict sperm retrieval and improve sperm retrieval rates; however, the evidence is insufficient. Further evidence accumulation is needed for salvage treatment in cases of failed sperm retrieval. In Japan, there is inadequate provision on the right to know the origin of children born from artificial insemination of donated sperm and the rights of sperm donors, as well as information on unrelated family members, and the development of these systems is challenging. In the future, it is hoped that the pathogenesis of nonobstructive azoospermia with an unknown cause will be elucidated and that technology for omics technologies, human spermatogenesis using pluripotent cells, and organ culture methods will be developed.
Topics: Child; Humans; Male; Azoospermia; Varicocele; Microdissection; Semen; Retrospective Studies; Gonadotropins; Testis
PubMed: 37737473
DOI: 10.1111/iju.15301 -
The Urologic Clinics of North America May 2020From a fertility perspective, men with azoospermia represent a challenging patient population. When no mature spermatozoa are obtained during a testicular sperm... (Review)
Review
From a fertility perspective, men with azoospermia represent a challenging patient population. When no mature spermatozoa are obtained during a testicular sperm extraction, patients are often left with limited options, such as adoption or the use of donor sperm. However, it has been reported that round spermatids can be successfully injected into human oocytes and used as an alternative to mature spermatozoa. This technique is known as round spermatid injection (ROSI). Despite the limitations of ROSI and diminished clinical success rates, the use of round spermatids for fertilization may have potential as a treatment modality for men with azoospermia.
Topics: Azoospermia; Female; Fertilization in Vitro; Humans; Infertility, Male; Male; Microdissection; Pregnancy; Pregnancy Outcome; Semen Analysis; Sperm Injections, Intracytoplasmic; Sperm Retrieval; Spermatids; Spermatogenesis
PubMed: 32272989
DOI: 10.1016/j.ucl.2019.12.004 -
Journal of the American Society of... May 2021Membranous nephropathy (MN) results from deposition of antigen-antibody complexes along the glomerular basement membrane (GBM). PLA2R, THSD7A, NELL1, and SEMA3B account...
BACKGROUND
Membranous nephropathy (MN) results from deposition of antigen-antibody complexes along the glomerular basement membrane (GBM). PLA2R, THSD7A, NELL1, and SEMA3B account for 80%-90% of target antigens in MN.
METHODS
We performed laser microdissection and mass spectrometry (MS/MS) in kidney biopsies from 135 individuals with PLA2R-negative MN, and used immunohistochemistry/immunofluorescence and confocal microscopy to confirm the MS/MS finding, detect additional cases, and localize the novel protein. We also performed MS/MS and immunohistochemistry on 116 controls and used immunofluorescence microscopy to screen biopsy samples from two validation cohorts. Western blot and elution studies were performed to detect antibodies in serum and biopsy tissue.
RESULTS
MS/MS studies detected a unique protein, protocadherin 7 (PCDH7), in glomeruli of ten (5.7%) PLA2R-negative MN cases, which also were negative for PLA2R, THSD7A, EXT1/EXT2, NELL1, and SEMA3B. Spectral counts ranged from six to 24 (average 13.2 [SD 6.6]). MS/MS did not detect PCDH7 in controls (which included 28 PLA2R-positive cases). In all ten PCDH7-positive cases, immunohistochemistry showed bright granular staining along the GBM, which was absent in the remaining cases of PLA2R-negative MN and control cases. Four of 69 (5.8%) cases in the validation cohorts (all of which were negative for PLA2R, THSD7A, EXT1, NELL1, and SEMA3B) were PCDH7-positive MN. Kidney biopsy showed minimal complement deposition in 12 of the 14 PCDH7-associated cases. Confocal microscopy showed colocalization of PCDH7 and IgG along the GBM. Western blot analysis using sera from six patients showed antibodies to nonreduced PCDH7. Elution of IgG from frozen tissue of PCDH7-associated MN showed reactivity against PCDH7.
CONCLUSIONS
MN associated with the protocadherin PCDH7 appears to be a distinct, previously unidentified type of MN.
Topics: Adult; Aged; Cadherins; Case-Control Studies; Cohort Studies; Female; Glomerulonephritis, Membranous; Humans; Laser Capture Microdissection; Male; Mass Spectrometry; Microscopy, Confocal; Middle Aged; Protocadherins
PubMed: 33833079
DOI: 10.1681/ASN.2020081165 -
Infection, Genetics and Evolution :... Jun 2020Single-cell sequencing (SCS) is a next-generation sequencing method that is mainly used to analyze differences in genetic and protein information between cells, to... (Review)
Review
Single-cell sequencing (SCS) is a next-generation sequencing method that is mainly used to analyze differences in genetic and protein information between cells, to obtain genetic information on microorganisms that are difficult to cultivate at a single-cell level and to better understand their specific roles in the microenvironment. By sequencing the whole genome, transcriptome and epigenome of a single cell, the complex heterogeneous mechanisms involved in disease occurrence and progression can be revealed, further improving disease diagnosis, prognosis prediction and monitoring of the therapeutic effects of drugs. In this study, we mainly summarized the methods and application fields of SCS, which may provide potential references for its future clinical applications, including the analysis of embryonic and organ development, the immune system, cancer progression, and parasitic and infectious diseases as well as stem cell research, antibody screening, and therapeutic research and development.
Topics: Epigenesis, Genetic; Genome; Genomics; High-Throughput Nucleotide Sequencing; Humans; Immunomagnetic Separation; Microdissection; Microfluidics; Organ Specificity; Sequence Analysis, DNA; Single-Cell Analysis; Species Specificity; Transcriptome
PubMed: 31958516
DOI: 10.1016/j.meegid.2020.104198 -
Fertility and Sterility Jun 2024
Topics: Humans; Male; Testis; Testicular Neoplasms; Microdissection
PubMed: 38554764
DOI: 10.1016/j.fertnstert.2024.03.023 -
The Analyst Jun 2021The human body comprises rich populations of cells, which are arranged into tissues and organs with diverse functionalities. These cells exhibit a broad spectrum of... (Review)
Review
The human body comprises rich populations of cells, which are arranged into tissues and organs with diverse functionalities. These cells exhibit a broad spectrum of phenotypes and are often organized as a heterogeneous but sophisticatedly regulated ecosystem - tissue microenvironment, inside which every cell interacts with and is reciprocally influenced by its surroundings through its life span. Therefore, it is critical to comprehensively explore the cellular machinery and biological processes in the tissue microenvironment, which is best exemplified by the tumor microenvironment (TME). The past decade has seen increasing advances in the field of spatial proteomics, the main purpose of which is to characterize the abundance and spatial distribution of proteins and their post-translational modifications in the microenvironment of diseased tissues. Herein, we outline the achievements and remaining challenges of mass spectrometry-based tissue spatial proteomics. Exciting technology developments along with important biomedical applications of spatial proteomics are highlighted. In detail, we focus on high-quality resources built by scalpel macrodissection-based region-resolved proteomics, method development of sensitive sample preparation for laser microdissection-based spatial proteomics, and antibody recognition-based multiplexed tissue imaging. In the end, critical issues and potential future directions for spatial proteomics are also discussed.
Topics: Ecosystem; Humans; Laser Capture Microdissection; Mass Spectrometry; Proteins; Proteomics
PubMed: 34042124
DOI: 10.1039/d1an00472g