Geneticky podmíněné faktory progrese vybraných chronických nefropatií.

Polycystická choroba ledvin je závažné geneticky podmíněné onemocnění vyskytující se u dospělých i dětských pacientů. Základní charakteristikou tohoto onemocnění je vznik a postupné zvětšování renálních cyst, které nahrazují funkční tkáň ledvin. U řady pacientů tak dochází k renálnímu selhání. Renální cysty se ovšem mohou vyskytovat i u řady dalších onemocnění, včetně multisystémových syndromů. To u některých pacientů komplikuje diferenciální diagnostiku onemocnění. V naší studii jsme se nejprve soustředili na diagnostiku a charakterizaci genotypově-fenotypových souvislostí u pacientů s polycystickou chorobou vznikající v dětském věku, později jsme naši studii rozšířili i na dospělé pacienty a pacienty s nejasnou klinickou diagnózou. Zároveň jsme zvětšili portfolio analyzovaných onemocnění, a to na řadu nemocí, u nichž se může vyskytnout fenotyp polycystických ledvin, i na onemocnění necystická. Během našeho projektu jsme metodou masivního paralelního sekvenování analyzovali 149 pacientů - 128 s cystickými a 21 s necystickými klinicky diagnostikovanými nefropatiemi. Zároveň byly nálezy ověřeny Sangerovou sekvenací u 176 příbuzných našich probandů. Mutační detekce dosahovala 59% u cystických pacientů a 43% u necystických pacientů. U řady pacientů molekulárně genetická analýza odhalila jinou...Polycystic kidney disease is a severe genetic disease occurring in both adult and pediatric patients. The basic characteristic of this disease is the development and progressive enlargement of renal cysts gradually replacing functional kidney tissue. This leads to renal failure in many patients. However, renal cysts may also occur in a number of other diseases, including multisystem syndromes. This complicates differential diagnosis in some patients. In our study, we first focused on the diagnosis and characterization of genotypic-phenotypic relationships in patients with polycystic disease arising in childhood, later we extended our study to adult patients and patients with unclear clinical diagnosis. At the same time, we expanded the portfolio of analyzed disorders to a number of diseases in which the phenotype of polycystic kidneys may occur, and noncystic diseases as well. During our project, massive parallel sequencing was used to analyze 149 patients - 128 with cystic and 21 with noncystic clinically diagnosed nephropathies. At the same time, the findings were verified by Sanger sequencing in 176 relatives of our probands. Mutation detection reached 59% in cystic patients, and 43% in non-cystic patients, respectively. In many patients, molecular genetic analysis revealed a different etiology...Department of Nephrology First Faculty of Medicine and General University HospitalKlinika nefrologie 1. LF UK a VFN1. lékařská fakultaFirst Faculty of Medicin

Mutational analysis of the TRPC6 gene in patients with nephrotic syndrome

Focal segmental glomerulosclerosis is one of the commonest cause of the nephrotic syndrome in adults patients. It is a damage of glomerulus characterized by leakage of proteins to urine and oedemas which usually develops into the end-stage renal disease within 10 years. Recently have been described familial forms of this disease which arise from injury to proteins making up filtration barrier of kidney. In 2005 non-selective ion channel TRPC6 was assigned among these proteins. In this thesis I focused on summarizing existing knowledge of the nephrotic syndrome, focal segmental glomerulosclerosis and involvement of TRPC6 in their origin. Second part of this work is devoted to the screening analysis of TRPC6 gene to discover possible mutations and polymorfisms in 47 patients with histologically proven focal segmental glomerulosclerosis or minimal change disease. The used methods were high resolution melting and direct sequencing. In the group of patients was detected no pathogenic mutation, only 2 known polymorfisms P15S and A404V and few changes which do not result in alteration of amino acid. So it seems TRPC6 gene mutations are a rare cause of the focal segmental glomerulosclerosis in adult patients in the Czech Republic

Mutational analysis of the TRPC6 gene in patients with nephrotic syndrome

Fokální segmentální glomeruloskleróza je jednou z nejčastějších příčin vzniku nefrotického syndromu u dospělých pacientů. Jedná se o poškození glomerulů charakterizované únikem proteinů do moči a edémy, jež se zpravidla do deseti let vyvine v chronické selhání ledvin. V posledních letech byla popsána familiární forma tohoto onemocnění vznikající po poškození proteinů tvořících filtrační bariéru ledvin. Mezi tyto proteiny byl roku 2005 přiřazen i neselektivní iontový kanál TRPC6. V této práci jsem se zaměřila na shrnutí dosavadních poznatků o nefrotickém syndromu, fokální segmentální glomeruloskleróze a zapojení TRPC6 na jejich vzniku. V praktické části jsem se zabývala screeningem mutací a polymorfismů v genu TRPC6 u 47 pacientů s histologicky potvrzenou fokální segmentální glomerulosklerózou či minimálními změnami glomerulů pomocí metody high resolution melting a přímou sekvenací. V souboru pacientů jsem neobjevila žádnou patogenní mutaci, pouze 2 známé polymorfismy P15S a A404V a několik záměn neměnících aminokyselinu. Zdá se tedy, že mutace v genu TRPC6 způsobující fokální segmentální glomerulosklerózu jsou v české populaci dospělých pacientů vzácné.Focal segmental glomerulosclerosis is one of the commonest cause of the nephrotic syndrome in adults patients. It is a damage of glomerulus characterized by leakage of proteins to urine and oedemas which usually develops into the end-stage renal disease within 10 years. Recently have been described familial forms of this disease which arise from injury to proteins making up filtration barrier of kidney. In 2005 non-selective ion channel TRPC6 was assigned among these proteins. In this thesis I focused on summarizing existing knowledge of the nephrotic syndrome, focal segmental glomerulosclerosis and involvement of TRPC6 in their origin. Second part of this work is devoted to the screening analysis of TRPC6 gene to discover possible mutations and polymorfisms in 47 patients with histologically proven focal segmental glomerulosclerosis or minimal change disease. The used methods were high resolution melting and direct sequencing. In the group of patients was detected no pathogenic mutation, only 2 known polymorfisms P15S and A404V and few changes which do not result in alteration of amino acid. So it seems TRPC6 gene mutations are a rare cause of the focal segmental glomerulosclerosis in adult patients in the Czech Republic.Katedra genetiky a mikrobiologieDepartment of Genetics and MicrobiologyPřírodovědecká fakultaFaculty of Scienc

Genetically determined progression factors of selected chronic nephropathies

Polycystic kidney disease is a severe genetic disease occurring in both adult and pediatric patients. The basic characteristic of this disease is the development and progressive enlargement of renal cysts gradually replacing functional kidney tissue. This leads to renal failure in many patients. However, renal cysts may also occur in a number of other diseases, including multisystem syndromes. This complicates differential diagnosis in some patients. In our study, we first focused on the diagnosis and characterization of genotypic-phenotypic relationships in patients with polycystic disease arising in childhood, later we extended our study to adult patients and patients with unclear clinical diagnosis. At the same time, we expanded the portfolio of analyzed disorders to a number of diseases in which the phenotype of polycystic kidneys may occur, and noncystic diseases as well. During our project, massive parallel sequencing was used to analyze 149 patients - 128 with cystic and 21 with noncystic clinically diagnosed nephropathies. At the same time, the findings were verified by Sanger sequencing in 176 relatives of our probands. Mutation detection reached 59% in cystic patients, and 43% in non-cystic patients, respectively. In many patients, molecular genetic analysis revealed a different etiology..

Focal segmental glomerulosclerosis and gene NPHS2

Katedra genetiky a mikrobiologieDepartment of Genetics and MicrobiologyFaculty of SciencePřírodovědecká fakult

Genetically determined progression factors of selected chronic nephropathies

Polycystic kidney disease is a severe genetic disease occurring in both adult and pediatric patients. The basic characteristic of this disease is the development and progressive enlargement of renal cysts gradually replacing functional kidney tissue. This leads to renal failure in many patients. However, renal cysts may also occur in a number of other diseases, including multisystem syndromes. This complicates differential diagnosis in some patients. In our study, we first focused on the diagnosis and characterization of genotypic-phenotypic relationships in patients with polycystic disease arising in childhood, later we extended our study to adult patients and patients with unclear clinical diagnosis. At the same time, we expanded the portfolio of analyzed disorders to a number of diseases in which the phenotype of polycystic kidneys may occur, and noncystic diseases as well. During our project, massive parallel sequencing was used to analyze 149 patients - 128 with cystic and 21 with noncystic clinically diagnosed nephropathies. At the same time, the findings were verified by Sanger sequencing in 176 relatives of our probands. Mutation detection reached 59% in cystic patients, and 43% in non-cystic patients, respectively. In many patients, molecular genetic analysis revealed a different etiology..

Mutational analysis of the TRPC6 gene in patients with nephrotic syndrome

Focal segmental glomerulosclerosis is one of the commonest cause of the nephrotic syndrome in adults patients. It is a damage of glomerulus characterized by leakage of proteins to urine and oedemas which usually develops into the end-stage renal disease within 10 years. Recently have been described familial forms of this disease which arise from injury to proteins making up filtration barrier of kidney. In 2005 non-selective ion channel TRPC6 was assigned among these proteins. In this thesis I focused on summarizing existing knowledge of the nephrotic syndrome, focal segmental glomerulosclerosis and involvement of TRPC6 in their origin. Second part of this work is devoted to the screening analysis of TRPC6 gene to discover possible mutations and polymorfisms in 47 patients with histologically proven focal segmental glomerulosclerosis or minimal change disease. The used methods were high resolution melting and direct sequencing. In the group of patients was detected no pathogenic mutation, only 2 known polymorfisms P15S and A404V and few changes which do not result in alteration of amino acid. So it seems TRPC6 gene mutations are a rare cause of the focal segmental glomerulosclerosis in adult patients in the Czech Republic