Grant proposal 1 Unraveling the molecular mechanisms involved in the therapeutic role of Tylophorinidine on behavioral alterations in Animal models of Autism Preclinical Approach Principal Investigator ID: 799357
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Slide1
Grant Proposal – DST/CSRI
Grant proposal
1
Slide2Unraveling the molecular mechanisms involved in the therapeutic role of Tylophorinidine on behavioral alterations in Animal models of Autism – Preclinical Approach
Principal Investigator
Dr.
Shyamjith
ManikkothAssociate ProfessorDepartment of PharmacologyYenepoya Medical College
Grant proposal
2
Slide3Contents
Rationale ObjectivesHypothesis
Methodology
Expected outcome
Timeline and Budget References Grant Proposal
3
Slide4Grant Proposal
4
AUTISM
Slide5Autism and Behavioral alterations
Grant proposal
5
Neurodevelopmental disorder in which the central nervous system are impaired.
A stereotype pattern of behaviors characterized by severe irregularity in communication, social awareness and skillfulness are commonly observed[1]
Has two main symptoms:
societal communication deficit
recurring or limited behaviors or interests.
Slide6Autism and Behavioral alterations
Grant proposal
6
Individuals with autism regularly show various allied symptoms
[2]
such as – Anxiety Self-injury Depression Epilepsy
Sleeplessness Attention deficits
Gastric disorders Hyperactivity
Irritability and aggression Mental retardation
Hypo-sensory problems Hyper-sensory problems
Slide7Autism and Behavioral alterations
Grant proposal
7
A variety of factors are implicated in the
etio
-pathogenesis of autism or autism spectrum disorder (ASD): [3]Weakened immune responses
Neuro
-inflammation
Unusual neurotransmission
Oxidative stress
Mitochondrial dysfunction
Environmental toxins
Epigenetic modifications
Slide8Need for the Study
Despite advances in early diagnosis and intercession, no therapy has been yet proven to completely reverse the core symptoms of autism. In the modern day therapy, there is no effective curative treatment for autism. This has, thus, driven the health care professionals to develop a new agent which can alleviate autism with less side effects
.
8
Grant proposal
Slide9In this perspective, role of phytochemicals comes into picture.
Considerable importance given to the active plant metabolites with minimal adverse effects to treat autism and behavioral alterations.
Grant proposal
9
Slide10Grant proposal
10
- Slender perennial climber
- Family-
Asclepiadaceae
- Indian
Ipecacuanha
-
Traditional uses:
bronchial asthma, rheumatism, allergies,
diarrhoea
Tylophora
indica
Slide11Grant proposal
11
Tylophora
indica
Previous studies
Anti- asthmatic, anti- inflammatory, anti anxiety, anti-rheumatic, analgesic, anticonvulsant, hepatoprotective and antioxidant activity.
Slide12Grant proposal
12
Tylophora
indica
From the HPLC-LCMS analysis it was found that Tylophorinidine is the active constituent present in the ethanolic extract of
Tylophora indica
and is responsible for many pharmacological properties. This phytoconstituent is an
isoquinoline
derivative.
Slide13Grant proposal
13
Phytochemical
Analysis Of TIEE using HPLC- LCMS
Major
Peaks
with
their
Retention
time
in
minutes
Peak
Area %
Molecular
Weight Observed
Possible
compound
5.79
39.325
360.0
Tylophorinidine
Slide14Objectives
To study the effect of TylophorinidineOn behavioural
abnormalities in Valproic acid induced autism in Wistar Albino rats.
On
neuroinflammation seen in autism by estimating IL-6, TNF-α and BDNF levels .On Neurotransmitters and Antioxidant levelsTo study the protective role of tylophorinidine in autism by quantifying the JAK - STAT pathway. To analyze whether there is any alteration in the methylation pattern in Promoter
CpG Islands of GABRB3.
Grant proposal
14
Slide15Hypothesis
Grant proposal
15
Having shown promising neuroprotective activity in various studies,
Tylophoridine
can play a pivotal role in autism treatment.
Slide16Materials and methods
16
WISTAR ALBINO RATS
TYLOPHORA INDICA
VALPROIC ACID
TYLOPHORINIDINE
Grant proposal
Slide17Sl.No
.
Work Plan
Duration
required
1.
Isolation
of tylophorinidine from
Tylophora indica
6 months
2
.
Studies on the therapeutic role of tylophorinidine on
behavioral
abnormalities in Valproic acid induced autism in
Wistar
albino rats
8 months
3
.
Estimation
of IL-6 , cytokines and BDNF levels to study the protective role of tylophorinidine in autism
3 months
4
.
Quantification
of neurotransmitter release in the brains of
autistic Wistar
rats
1 month
5
.
Evaluating
the protective role of tylophorinidine on oxidative stress induced in autistic animals
1 month
6
.
To
assess the role of tylophorinidine in molecular alterations associated with JAK-STAT pathway
3 months
7
.
To analyze whether there is any alteration in the
methylation
pattern in Promoter
CpG
Islands of
GABRB3
8 months
17
Grant proposal
Slide18Methodology
Behavioral Abnormalities - Rota – rod test, touch response and pain response -
Actophotometer
test and Open field test
- Elevated plus maze and light dark arena tests - Morris water maze and runway apparatus test - Forced swimming test and tail suspension testBiochemical Estimations -
ELISA kits - Immunofluorescence - Western Blotting technique
18
Grant proposal
Slide193. Neurotransmitter and Antioxidant Investigations
- Spectrophotometric and Spectrofluorimetric techniques
- Flow
cytometric
assays4. JAK – STAT pathway detection - ELISA kits5. DNA alteration -
qRT – PCR 19
Methodology
Grant proposal
Slide20Expected outcome and Applicability
The study aims at understanding and unraveling the molecular mechanisms associated with the therapeutic potential of tylophorinidine in autism using experimental models.
Results of the study will pave an insight into the possibility of developing tylophorinidine as a novel sole therapeutic agent for mitigating behavioral abnormalities in autism.
20
Grant proposal
Slide2121
Grant proposal
Activities
Time schedule (Months)
1-6
7-12
13-18
19-24
25-30
31-36
Literature survey
Isolation of tylophorinidine from
Tylophora indica
1
2
3
4
5
6
Evaluation of therapeutic potential of tylophorinidine on autism model
7
8
9
13
14
10
11
12
Estimation of IL-6 , cytokines and BDNF levels
15
16
17
Quantification of neurotransmitter release
18
Protective role of tylophorinidine on oxidative stress in autism
19
Assess the role of tylophorinidine in molecular alterations associated with JAK-STAT pathway
20
21
22
Analysis of alteration in DNA
methylation
25
26
27
23
24
28
29
30
Data compilation, publications
31
32
33
34
35
36
Slide22Budget
22
Grant proposal
ITEMS
BUDGET
Total (INR)
1st Year
2nd Year
3rd Year
A.
Recurring
Salaries/wages
1,92,000
1,92,000
1,92,000
5,76,000
Consumables
10,00,000
9,00,000
8,00,000
27,00,000
Travel
50,000
50,000
50,000
1,50,000
Other costs
20,000
20,000
20,000
60,000
B.
Equipment
10,00,000
10,00,000
Grand total (A+B
) Total FEC*
44,86,000
Slide2323
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Grant proposal
Slide24THANK YOU
24
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Slide2525
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Grant proposal