Transcript No Slide Title

PATENT PROJECT FINAL PRESENTATION
TITLE :
USE OF PROSTAGLANDIN E 2 RECEPTOR
GENE(EP2) FOR RELAXATION OF CILIARY
MUSCLES AND THEREBY HELP IN THE
TREATMENT OF FUNCTIONAL MYOPIA
-SACHIN JAMBAWALIKAR
ABSTRACT :
A NOVEL APPROACH TO USE GENE THERAPY TO
TRANSFECT THE CILIARY MUSCLE CELLS WITH
PROSTAGLANDIN 2 RECEPTOR GENE.THE EXPRESSION OF
THIS GENE WILL INDUCE A SERIES OF EVENTS WHICH
INCLUDE THE CYCLIC -AMP(SECOND MESSENGER)PATHWAY
AND CAUSE THE RELAXATION OF THE CILIARY MUSCLE IN
THE FUNCTIONAL MYOPIC PATIENTS.
BACKGROUND:
MYOPIA: ITS DEFINED AS THE STATE WHERE THE RAYS
OF LIGHT EMERGING FROM A DISTANT OBJECT FOCUS AT
A POINT IN FRONT OF THE RETINA THEREBY PRODUCING
A BLURRED IMAGE ON THE RETINA .
TYPES:
STRUCTURAL MYOPIA: OVER ELONGATION OF THE EYE
BALL CAUSES STRUCTURAL MYOPIA
FUNCTIONAL MYOPIA:
CILIARY MUSCLES ARE WEAKENED AND COMPLETE
RELAXATION DOESNOT TAKE PLACE THEREBY MAKING
THE LENS MORE CURVED .
AVAILABLE TREATMENT OPTIONS :
1) OPTICAL CORRECTION
2) MEDICAL(PHARMACEUTICAL): CYCLOPLEGIC AGENTS
3) VISION THERAPY
4) ORTHOKERATOLOGY
5) REFRACTIVE SURGERY
6) FOOD AND NEUTRITION THERAPY
HYPOTHESIS AND ASSUMPTION ON WHICH THIS
INVENTION IS BASED.
THE FUNCTIONAL MYOPIA WHEN SEEN FROM
THE MOLECULAR LEVEL MIGHT BE CAUSED DUE TO A
PROBLEM IN THE CELL SIGNALLING MECHANISM.
IT MIGHT BE CAUSED DUE TO DEGRADATION OR
PERMANENT BLOCKING OF RECEPTORS SUCH AS
PROSTAGLANDIN 2E ALPHA RECEPTOR. A RESULT OF
THIS IS INCOMPLETE RELAXATION OF CILIARY MUSCLE
THIS CAUSES THE LENS TO REMAIN BULGED DUE TO
IMPROPER ACCOMODATION AND THE PERSON
BECOMES MYOPIC.
DETAILED EXPLAINATION OF THE INVENTION:
1) VIRUS VECTOR USED FOR GENE DELIVERY.
2)GENE DELIVERY PROCEDURE
3)PGE2 AND EP2 RECEPTOR
4)PGE2 INITIATED CYCLIC AMP PATHWAY AND
RELAXATION OF CILIARY MUSCLE
VIRUS VECTOR :
-THE VIRUS VECTOR USED WAS HSV-1,HrR3.
-THE VIRUS GENETIC SEQUENCE WAS ALTERED BY
ADDITION OF EP2 RECEPTOR GENE.
-HSV1 VIRUS IS USED AS COMPARED TO ADENOVIRUS
AS HSV1 HAS HIGH EFFICIENCY IN GENE DELIVERY TO
THE NERVOUS SYSTEM. COMPARED TO ADENO VIRUS
-HSV1 IS KNOWN TO HAVE MUCH HIGHER CAPACITY
FOR ACCOMODATING MULTIPLE THERAUPATIC GENES.
GENE DELIVERY:
-A 30 GAUGE NEEDLE WAS THREADED THROUGH THE
TEMPORAL CORNEA OF ANASTHETIZED PATIENT.
-THE EP2 RECEPTOR GENE WITH ITS VECTOR IS
INJECTED IN THE ANTERIOR CHAMBER.
-THE HSV-1 VIRUS TRANSFECTS THE GENE INTO THE
MUSCLE CELLS OF THE CILIARY BODY.
PGE2-ALPHA AND EP2 RECEPTORS :
-PROSTAGLANDINS ARE A GROUP OF EICOSANOID.
-THESE SIGNALLING MOLECULES OFTEN ACT IN AN
AUTOCRINE MODE IN MATURE MAMMALS.
-THEY ARE FATTYACID DERIVATIVES MADE BY CELLS IN
ALL MAMMALIAN TISSUE .
-THEY ARE MADE FROM PRECURSORS(MAINLY
ARACHIDONIC ACID)WITH 20CARBON FATTY ACID CHAINS
AND ATLEAST THREE DOUBLE BONDS.
-PROSTAGLANDIN ARE A PRODUCT OF
CYCLOOXYGENASE -CATALYSED METABOLISM OF
ARACHIDONIC ACID.
-EP2 RECEPTOR IS A G-PROTIEN LINKED RECEPTOR.
-IT IS A TRANSMEMBRANE PROTEIN WITH 7 PASSES
THROUGH THE PLASMA MEMBRANE.
-THE EP2 RECEPTOR ACTS INDIRECTLY TO REGULATE
THE ACTIVITY OF A SEPARATE PLASMA MEMBRANE
BOUND TARGET PROTEIN WHICH IS THE CALCIUM ION
CHANNEL .
PGE2 INTIATED CYCLIC AMP PATHWAY AND
RELAXATION OF CILIARY MUSCLE
STEPS:
1) THE EP2 RECEPTOR GENE ON EXPRESSION PRODUCES
THE EP2 RECEPTOR PROTEIN.
2) THIS PROTEIN WILL MOVE TOWARDS THE PLASMA
MEMBRANE AND WILL FORM A SEVEN PASS TRANS
MEMBRANE PROTEIN.
3)WHEN THE LIGAND (PGE)RELEASED BY
NEIGHBOURING CELL ATTACHES TO EP2 RECEPTOR IT
CAUSES A CONFORMATIONAL CHANGE IN THE SUBUNIT OF THE G PROTEIN.
4)THIS WILL SIGNAL ADENYLYL CYCLASE(AC) A
MEMBRANE BOUND INTRACELLULAR ENZYME WHICH
CATLYZES THE CONVERSION OF ATP TO CYCLIC AMP.
5) PRODUCTION OF CYCLIC AMP WILL ACTIVATE CYCLIC
AMP DEPENDENT PROTEI KINASE(PKA).
6)PKA WILL PHOSPHORYLATE SEVERAL DIFERENT
INTRACELLULAR PROTEINS THAT ONCE ACTIVATED
LEAD TO MUSCLE RELAXATION BY OPENING OF THE
CALCIUM CHANNEL AND A SERIES OF OTHER EVENTS
THERE BY CAUSING COMPLETE REPOLARIZATION OF THE
CELL.
CLAIMS :
1 )EP2 ALPHA RECEPTOR GENE IS TRANSFECTED INTO THE
CILIARY MUSCLE CELLS.THE EXPRESSION OF THIS GENE
WILL CAUSE RELEASE OF PROTEIN WHICH ACTS AS A G
PROTEIN LINKED RECEPTORAND ON INTERACTION WITH
PGE2 LIGAND HELPS IN RELAXATION OF THE CILIARY
MUSCLE BY GOING THROUGH THE CYCLIC AMP
PATHWAY.THIS CAN BE USED FOR THE TREATMENT OF
FUNCTIONAL MYOPIA.
2)THE METHOD OF CLAIM 1 WHERE IN THE VIRUS VECTOR
CAN BE RETROVIRUS,ADENOVIRUS,ADENOASSOCIATED
VIRUS(AAV),VACCINIA VIRUS AND POLIO VIRUS.
3)THE METHOD IN CLAIM 1 WHERE IN THE ARE OF
TREATMENT IS GLAUCOMA.
4) THE METHOD OF CLAIM 1 WHERE THE RECEPTOR GENE
CAN BE PROSTAGLANDIN 2FALPHA,2A ALPHA,2B
ALPHA,2C ALPHA AND 2D ALPHA.
5)THE METHOD OF CLAIM 1 WHERE IN THE TRANS
FECTED GENE IS ADRENERGIC BETA2 RECEPTOR GENE .
6)NITRIC OXIDE SYNTHASE GENE CAN ALSO BE
DELIVERED BY THE METHOD 1.THIS WILL PRODUCE
NITRIC OXIDE .NO WILL PASS THROUGH THE PLASMA
MEMBRANE OF THE TARGET CELLS AND ACTIVATE
INTRACELLULAR ENZYMES USUALLY GUANYLYL
CYCLASE WHICH WILL PRODUCE CYCLIC GMP. THIS IN
TURN PRODUCES PKG WHICH PRODUCES RELAXATION AS
IN THE CYCLIC AMP PATHWAY.