Using Sperm Transcriptome in Search for Novel Biomarkers of Male Infertility

Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically useful tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of...S problémem neplodnosti se v současně době setkává 10 až 15 procent párů. Přitom přibližně v 50 procentech případů se na problému podílí mužský faktor. Většina příčin mužské neplodnosti zůstává neznámá. Proto je velmi aktuální otázka nalezení jednoduchých a v klinické praxi použitelných nástrojů diagnostiky mužské neplodnosti. V této práci jsme se na základě analýzy transkriptomu spermií pokusili najít geny, u nichž by mohl rozdíl v expresi mezi normálními a patologickými spermiemi poskytnout informace o molekulární podstatě patologie. Stanovení exprese takových genů by mohlo sloužit pro neinvazivní diagnostiku mužské infertility. Do této studie bylo zahrnuto 67 vzorků spermií včetně 16 kontrolních od zdravých mužů. Vzorky neplodných pacientů byly rozděleny do tří skupin podle morfologie a motility na základě údajů spermiogramu. Do skupiny astenoteratozoospermie bylo zařazeno 20 vzorků vykazujících sníženou motilitu a abnormální morfologii, do skupiny astenozoospermie bylo zařazeno 15 vzorků spermií s normální morfologií, ale sníženou motilitou, do třetí skupiny, normospermie, bylo zařazeno 16 vzorků neplodných pacientů s normálním spermiogramem z hlediska počtu spermií, motility i morfologie. Dvacet čtyři vzorků ze všech čtyř skupin (tři skupiny infertilních mužů a jedna skupina kontrolní - sedm...Katedra buněčné biologieDepartment of Cell BiologyPřírodovědecká fakultaFaculty of Scienc

Using Sperm Transcriptome in Search for Novel Biomarkers of Male Infertility

Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically usefµl tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of...S problémem neplodnosti v současně době se setkává 10 až 15 procent párů. Přitom přibližně v 50 procentech případů se na problému podílí mužský faktor. Většina příčin mužské neplodnosti zůstává neznámá. Proto je velmi aktuální otázka nalezení jednoduchých a v klinické praxi použitelných nástrojů diagnostiky mužské neplodnosti. V této práci jsme se na základě analýzy transkriptomu spermií pokusili najít geny, u nichž by mohl rozdíl v expresi mezi normálními a patologickými spermiemi poskytnout informace o molekulární podstatě patologie. Stanovení exprese takových genů by mohlo sloužit pro neinvazivní diagnostiku mužské infertility. Do této studie bylo zahrnuto 67 vzorků spermií včetně 16 kontrolních od zdravých mužů. Vzorky neplodných pacientů byly rozděleny do tří skupin podle morfologie a motility na základě údajů spermiogramu. Do skupiny astenoteratozoospermie bylo zařazeno 20 vzorků vykazujících sníženou motilitu a abnormální morfologii, do skupiny astenozoospermie bylo zařazeno 15 vzorků spermií s normální morfologií, ale sníženou motilitou, do třetí skupiny, normospermie, bylo zařazeno 16 vzorků neplodných pacientů s normálním spermiogramem z hlediska počtu spermií, motility i morfologie. Dvacet čtyři vzorků ze všech čtyř skupin (tři skupiny infertilních mužů a jedna skupina kontrolní - sedm...Katedra buněčné biologieDepartment of Cell BiologyFaculty of SciencePřírodovědecká fakult

Using Sperm Transcriptome in Search for Novel Biomarkers of Male Infertility

Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically useful tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of..

Using Sperm Transcriptome in Search for Novel Biomarkers of Male Infertility

Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically usefµl tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of..

The problem of infertility and the methods of resolving..

4 Abstract This diploma thesis is concerned with modern methods of treatment for infertility. It examines to what extent and in what cases are these methods used and what risk does it pose for a woman's and a child's health. Paticular emphasis is placed on the methods of assisted reproductive technology as the most used, effective and the most invasive. Nowadays 10-15% of couples face the problem of infertility. In most of the cases this problem can be solved with the help of hormonal therapy, surgery or assisted reproductive technology. The most common complication of ART (assisted reproductive technology) is ovarian hyperstimulation syndrome which leads to higher frequency of miscarriages and can be fatal for a woman. Since more that one embryo is usually transferred in ART children conceived with the help of this method can face different problems related to multiple pregnancy such as higher risk of preterm birth, higher risk of congenital disorder or intrauterine growth retardation. Key words: ART, assisted reproductive technology, infertility, congenital disorder, multiple pregnanc

Somatic Mutations in Exon 7 of the <i>TP53</i> Gene in Index Colorectal Lesions Are Associated with the Early Occurrence of Metachronous Adenoma

(1) Background: this prospective study was focused on detailed analysis of the mutation heterogeneity in colorectal lesions removed during baseline (index) colonoscopy to identify patients at high risk of early occurrence of metachronous adenomas. (2) Methods: a total of 120 patients after endoscopic therapy of advanced colorectal neoplasia size ≥10 mm (index lesion) with subsequent surveillance colonoscopy after 10–18 months were included. In total, 143 index lesions and 84 synchronous lesions in paraffin blocks were divided into up to 30 samples. In each of them, the detection of somatic mutations in 11 hot spot gene loci was performed. Statistical analysis to correlate the mutation profiles and the degree of heterogeneity of the lesions with the risk of metachronous adenoma occurrence was undertaken. (3) Results: mutation in exon 7 of the TP53 gene found in the index lesion significantly correlated with the early occurrence of metachronous adenoma (log-rank test p = 0.003, hazard ratio 2.73, 95% confidence interval 1.14–6.56). We did not find an association between the risk of metachronous adenomas and other markers monitored. (4) Conclusions: the findings of this study could lead to an adjustment of existing recommendations for surveillance colonoscopies in a specific group of patients with mutations in exon 7 of the TP53 gene in an index lesion, where a shortening of surveillance interval may be warranted