1.
Three Dimensional Cerebral Cavernous
Malformation Organoids to Advance Drug Discovery
for CCM.

2.
- Vascular Endothelial Cells
- Endothelial Cells Junctions
- Cerebral Cavernous Malformation
- Spheroids & Organoids
- Methodology
Content

3.
Vascular Endothelial Cells
Single layer of cells
Formed from embryonic
mesoderm
Provide a barrier between the
blood and body’s tissues

4.
Endothelial Cells Junctions

5.
Cerebral Cavernous Malformation

6.
Cerebral Cavernous Malformations
- Disorder known as CCM, mostly affects the neurovasculature.
- Expanded caverns and thick capillaries are structured in a back-to-back
pattern.
- This defect lack all of the components needed for mature blood vessels,
such as smooth muscle and elastic tissue, and the presence of defective cell–
cell junctions.
- CCMs can also be found in the retina, liver, kidney, and on the skin

7.
- This malformation is found in 1 out of every 200–250 people.
- Many factors that can influence the symptoms, such as the location,
number, and size of lesions.
- The gold standard for CCM detection is 3T Magnetic Resonance Imaging.
- Patients are given various treatments to control or slow the progression of
their disease.
Cerebral Cavernous Malformations

8.
Cerebral Cavernous Malformations
1. Micro–computed tomography (CT)
analysis of CCM lesions in Ccm1
2. Magnetic resonance imaging of a patient
with a pathogenic CCM1 germline mutation
- This figure indicates a major CCM and
many smaller CCMs in both hemispheres
1
2

9.
CCM Pathogenesis
- Scientists have identified two types for this disorder:
* Sporadic causes, which occur in people without a CCM family history (80%)
* Familial cerebral cavernous malformation (fCCM) which are mostly
associated with the presence of a single or multiple lesions (20%)
- Three genes are responsible for two types of CCM :
* CCM1/KRIT1
* CCM2/MGC4607
*CCM3/PDCD10

10.
* CCM1/KRIT1
 Is expressed in the embryo and then gradually decreases; it is primarily detected
in the nervous and epithelial tissues.
 More than 300 mutations have been identified in this gene, such as frameshift,
nonsense, missense, and splice site sequence variants lead to early stop codons
and defects in proteins.
*CCM2/MGC4607
 Expressed in the endothelium of various organs, and many defects were revealed
in artery and vein formation when this gene was suppressed.

11.
* CCM3/PDCD10
 Have significant and severe symptoms.
 Highly conserved pro-apoptotic gene
with over 70 mutations that cause
protein dysfunction.
 When it is suppressed in animals, it
causes aberrant vasculogenesis and
hematopoiesis

12.
- More than 350 distinct CCM1/CCM2/CCM3 mutations cause sporadic or
fCCM.
- Animal experiments show that CCM gene knockout mice die during
embryogenesis due to heart and vascular defects.
- Mice that is carrying only one mutated allele rarely develop cavernous
malformations.
Cerebral Cavernous Malformations

13.
Spheroids
- Type of 2D monolayer that can be grown on cell culture plates or other platforms.
- Many characteristics such as self-renewal, formation of differentiated offspring,
and protein secretion, lead stem cells spheroids to be in tissue engineering and
regenerative medicine.

14.
Organoids
- Three-dimensional (3D) organ analogs that can be made from pluripotent stem
cells or adult stem/progenitor cells and cultured in vitro.
- Faithfully, albeit imperfectly, replicates the embryonic development and
physiology of primary tissues in vivo.

15.
- Many limitations found for these systems:
* Still lack numerous properties of normal human tissues, such as
connections with supporting cells.
* Absence of vascular circulation and immune systems in brain
organoids technique.
Organoids

16.
* The aim of our study to develop CCM spheroid to CCM
organoids which help others in explore proper treatment
and apply drugs on it.

17.
How to develop the CCM spheroids to organoids?

18.
Methodology
Isolated ECs
Matrigel

19.
Methodology