This document summarizes a study that investigated the role of macrophage-derived CCN3 in the progression of calcific aortic valve disease. The study used techniques like immunofuorescence, PCR, Western blot, and ELISA to analyze CCN3 levels and interactions with other proteins in normal and calcified aortic valves. Results found CCN3 and other proteins like CD68, BMP2, and Vimentin present in calcified valves. The conclusion is that molecular biology techniques have helped further understanding of pathologies and development of diagnoses and treatments.

1. Universidad Pontificia Bolivariana
III semester
Maria Fernanda Oliveros Goez

2. Calcifc aortic valve disease
The most prevalent form of heart
valve disease, is characterized by
the thickening of the aortic valve
due to signifcant fbrosis and/ or
calcifcation.
INTRODUCTION

3. CCN3 GENE
The protein encoded by this gene is a
small secreted cysteine-rich protein
and a member of the CCN family of
regulatory proteins. CNN family
proteins associate with the
extracellular matrix and play an
important role in cardiovascular and
skeletal development.
INTRODUCTION

4. GENERAL
OBJECTIVE
In the present study, was investigated
the role of macrophage-derived
CCN3 in the progression of calcifc
aortic valve disease.

5. METHODS
This technique allows the
detection of antigens from
microorganisms to which specific
fluorescein-labeled antibodies
have been bound using a
fluorescence microscope
This technique consists of the
detection of an antigen through
an antibody linked to an enzyme,
through some detectable product,
such as a change in color, which
would make it possible to
measure the antigen in the
sample using spectrophotometry.
ELISA IMMUNOFUORECENSE

6. METHODS
Before we start learning, let's
try to answer this question:
It is based on the DNA polymerase
chain reaction (PCR) where a
known piece of DNA is copied and
amplified billions of times.
Real-time is a modality of end-
point PCR, where the accumulation
of amplified DNA is detected and
quantified as the reaction
progresses.
The method involves the use of gel
electrophoresis to separate the
proteins in the sample. The
separated proteins are transferred
from the gel to the surface of a
membrane. The membrane is
exposed to a specific antibody
against the protein under study.
Antibody binding is detected using a
radioactive or chemical label
WESTERN BLOT PCR

7. RESULTS
A. Representative immunofuorescence
images of CCN3 and CD68 on either
normal or calcifed patient aortic
valves
C. Immunofuorescence of
BMP2 and Vimentin on calcifed
aortic valves.

9. AUTHORS STATEMENT
AGREE /
DISAGREE
Minamizato T, Sakamoto K,
Liu T, Kokubo H, Katsube K,
Perbal B, Naka‐
mura S, Yamaguchi A.
A prior in vitro study showed that CCN3 inhibits BMP2-induced osteoblast
differentiation by limiting BMP signaling and activation of the Notch pathway
Wu W, Hu X, Zhou X,
Klenotic PA, Zhou Q, Lin Z.
Exacerbation of NAFLD as a result of myeloid deficiency of CCN3 provided
additional evidence that myeloid CCN3 could impact the behavior of other cell
types via a paracrine manner
Kumar S, Kang DW,
Rezvan A, Jo H
Additionally, others have demonstrated a single injection of rAAV8/D377Y-
mPCSK9 coupled with HFD is a feasible approach to study cardiovascular
calcifcation
DISCUSSION

10. CONCLUSION
Molecular biology has generated a great impact
at the research level since it helps to have a
greater vision of many pathologies, contributing
to faster diagnoses and more effective
treatments.
The use of molecular biology has aided the study
of human genetics and the development,
diagnosis, and treatment of related diseases.