Manuela Amaya CossioManuela Zapata Arismendy.

Molecular BiologyIII Semester

ETHNIC DISPARITY IN 21-HYDROXYLASE GENE MUTATIONS

IDENTIFIED IN PAKISTANI CONGENITAL ADRENAL

HYPERPLASIA PATIENTS

Aysha H Khan1, Muniba Aban1, Jamal Raza2, Naeem ul Haq3, Abdul Jabbar3, Tariq Moatter1

INTRODUCTION

CONGENITAL ADRENAL HYPERPLASIA (CAH).

• Autosomal recessive disorder.

• Characterized by defects in any of the five enzymes required to sintheside cortisol, primarity of 21 hidroxylase enzyme (21OH).

• It deficiency results in reduced synthesis of cortisol and aldosterone.

CAH phenotypes vary whit the dregree of the enzyme deficiency:

• Complete Deficiency Enzyme: Prenatal virilization in newborns females and salt wasting (SW – CAH), in both sexes, in the neonatal period.

• The partial enzyme deficiency: Classic simple virilization (SV-CAH), presenting with prenatal virilization in females and pseudoprecosiaus puberty in both sexes.

INTRODUCTION

• Mild deficiency of 21OH: Causes non classical disease (NC – CAH) with pseudoprecocius puberty, hirsutism, acne, and reduce fertility at later age.

• Clinical Diversity in CAH is primarity due to mutations, CYP21A2, gene which encodes 21 OH.

INTRODUCTION

CYP21A2 GENE FUNCTIONS

• The CYP21A2 gene function is encode the 21-hidroxylase enzyme (21OH)

• It´s located within the human leukocyte antigen class III region on the short arm of cromosome 6.

• This has a highly homologous pseudo gene CYP21A1P.

• Both have 3-4 kb long and each gene consists of 10 exons. 95% of the CAH cases are caused by a specific point mutations an deletions in the CYP21A2 gene.

INTRODUCTION

RELATIONSHIP BETWEEN CAH AND

CYP21A2 GENE

• This disease is caused by CYP21A2 gene mutation, causing in the adrenal a partial or total decrease in the cortisol and aldostorone production, therefore in the patients is evidence a male phenotype in female neonate, or salt wasting in both sexes.

• The adrenal hyperplasia was classified by phenotypes that vary with the degree of enzime deficiency.

INTRODUCTION

Correlate genotype with the phenotype in a randomly selected

cohort of congenital adrenal hyperplasia patients, will facility and

promote prenatal diagnostic of disease, in the Pakistan population

throgh the knowledge of the mutations in CYP21A2 gene

GENERAL OBJECTIVE

POBLACIÓN DE ESTUDIO

Diagnostico de hiperplasia suprarrenal congénita.

Presentación Clínica

Evaluación Bioquímica

Investigación hormonal

SW (Derrochador

de sal)

SV(Virilizacion

simple)

NC (No clásica)

Nivel sodio

17 hidroxi - progesterona

Nivel de potasio

MATERIALES Y METODOS

•Falta de crecimiento Vómitos

•Hiperpotasemia Hiponatremia

•Azotemia Acidosis Metabólica

•Deshidratación.

CAH-SW•Genitales

Ambiguos en recién nacidas.

• Ampliación del pene

• Edad ósea avanzada

• Pubertad precoz.

CAH - SV

CAH – NC

MATERIALES Y METODOS

•Hirsutismo•Acné•Pubertad precoz

MATERIALES Y METODOS

• Muestras de sangre con EDTA.

• ADN genómico extraído de los leucocitos, con promega.

• ADN purificado fue proyectado por mutaciones en el gen CYP21A2 utilizando PCR- ARMS.

• PCR: En esta técnica de

laboratorio se tiene como objetivo

la amplificación directa de un gen

o de un fragmento de DNA, o

indirecta de RNA. Esta se basa en

la amplificación enzimática in vitro

la cual es el incremento

geométrico del numero de copias

de una secuencia particular del

DNA.

MATERIALES Y METODOS

PCR – ARMS: (sistema refractario de una amplificación) Se utiliza

dos cebadores idénticos en su secuencia excepto en su nucleótido

3'. Uno tiene el nucleótido 3' complementario al gen salvaje,

mientras que el otro cebador 3' es complementario al gen mutado.

La amplificación sólo ocurrirá con emparejamientos perfectos, con

lo que dependiendo de si el cebador (primer) origine una señal

positiva, podremos distinguir el gen salvaje del gen mutado, ARMS

es un método especialmente sensible para la detección de

mutaciones.

MATERIALES Y METODOS

RESULTADOS

RESULTADOS

RESULTADOS

Khan H Aysha -The health issues of gender ambiguity are not well studied in the Pakistan population-Illnesses like CAH are considered a family matter rather than an individual occurrence

-Tools, This is the need, because even when they know how to do it, world need to be better prepared for helping those important patients

R Bhanji and AH Khan

-children with CAH are usually brought to medical attention late in life

-This is part of the main problem this study is train to improve and make it better.

R Bhanji and A Thilen

-the median chronological age atthe time of diagnosis was comparatively high and only41% cases were diagnosed during the newborn period

-A way to improve the main problem, whit knowledge and tools

Pakistan faces genetic disasters and Pakistan: lack of healthcare acces.

-The management of genetic disorders has not received its due share in Pakistan, healthcera systemdoes not include genetic disorders as CAH

-Government help means the open door for getting a better livestyle.

DISCUSSION

CONCLUSIONS

1. The main important of this work, goes around person`s live,

because when parents have to take those hard decitions

science should be there helping them.

2. Wolrd is not prepared for persosn that need special atention, and

i think this arms-pcr examen should`ve been done lot of time,

giving patients a better live and psycological development.

3. In this study of congenital adrenal hyperplasia, it reported major

problems on births associated with this pathology, principally the

presence of sexual ambiguity, because this genetic defect may be

know before the birth, for improvement the quality  of life in this

patients.

CONCLUSIONS

4. The relationship between phenotype and genotype, gives a

clearer approach to congenital adrenal hyperplasia. This

disease is expressed with different signs and symptoms that

make the medical consultation will be directly aimed at a

specified classification, and according to a  diagnosis may be

hormone deficiency due to total or partial absence of the

enzyme.

CONCLUSIONS

BIBLIOGRAFIA

• Martínez. LM, Biología Molecular: Libro de clase, Sexta

Edición, Editorial Universidad Pontificia Bolivariana,

Medellín, 2011.

GRACIAS!