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Grant Proposal – DST/CSRI Grant Proposal – DST/CSRI

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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

grant autism 000 proposal autism grant proposal 000 tylophorinidine model kim role syndrome mecp2 behavioral disorders alterations spectrum references

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Slide1

Grant Proposal – DST/CSRI

Grant proposal

1

Slide2

Unraveling 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

Slide3

Contents

Rationale ObjectivesHypothesis

Methodology

Expected outcome

Timeline and Budget References Grant Proposal

3

Slide4

Grant Proposal

4

AUTISM

Slide5

Autism 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.

Slide6

Autism 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

Slide7

Autism 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

Slide8

Need 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

Slide9

In 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

Slide10

Grant proposal

10

- Slender perennial climber

- Family-

Asclepiadaceae

- Indian

Ipecacuanha

-

Traditional uses:

bronchial asthma, rheumatism, allergies,

diarrhoea

Tylophora

indica

Slide11

Grant proposal

11

Tylophora

indica

Previous studies

Anti- asthmatic, anti- inflammatory, anti anxiety, anti-rheumatic, analgesic, anticonvulsant, hepatoprotective and antioxidant activity.

Slide12

Grant 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.

Slide13

Grant 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

Slide14

Objectives

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

Slide15

Hypothesis

Grant proposal

15

Having shown promising neuroprotective activity in various studies,

Tylophoridine

can play a pivotal role in autism treatment.

Slide16

Materials and methods

16

WISTAR ALBINO RATS

TYLOPHORA INDICA

VALPROIC ACID

TYLOPHORINIDINE

Grant proposal

Slide17

Sl.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

Slide18

Methodology

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

Slide19

3. 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

Slide20

Expected 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

Slide21

21

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

Slide22

Budget

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

Slide23

23

References

Christina M.

Corsello

. Early Intervention in Autism. Infants & Young Children, 2005; 18(2): 74-85

Emily JH.

Jonesa

,

Teodora

Gligaa

, Rachael

Bedfordc

, Tony

Charmanb

, Mark H.

Johnsona

.

Developmental pathways to autism: A review of prospective studies of infants at risk. Neuroscience and

Biobehavioral

Reviews, 2014; 39:1–33

Necmi

Namala

,

Hayriye

Ertem

Vehitb

,

Selcuk

Koksalc

. Do autistic children have higher levels of caries? A cross-sectional study in Turkish children. J Indian Soc

Pedod

Prev

Dent, 2007;97-100

Muthu

MS,

Prathibha

KM. Management of a child with autism and severe

bruxism

: A case report. J Indian Soc

Pedod

Prevent Dent,2008: 82-84

Daniel A.

Rossignol

. Novel and emerging treatments for autism spectrum disorders: A systematic review. Annals of Clinical Psychiatry 2009;21(4):213-236.

Alexis

Tchaconasa

and Andrew

Adesman

. Autism spectrum disorders: a pediatric overview and update.

Curr

Opin

Pediatr

2013, 25:130–143

Helen V.

Ratajczak

. Theoretical aspects of autism: Causes—A review. Journal of

Immunotoxicology

, 2011; 8(1): 68–79

D.

Ebrahimi-Fakhari

, M.

Sahin

, Autism and the synapse: Emerging mechanisms and mechanism-based therapies.

Curr

.

Opin

. Neurol. 28, 91–102 (2015)

Grant proposal

Slide24

THANK YOU

24

Grant proposal

Slide25

25

R. J. Kelleher 3rd, M. F. Bear, The autistic neuron: Troubled translation? Cell 135, 401–406 (2008).

H. Y.

Zoghbi

, M. F. Bear, Synaptic dysfunction in neurodevelopmental

disorders associated with autism and intellectual disabilities. Cold Spring Harb.

Perspect

. Biol. 4, a009886 (2012).

T. C.

Südhof

,

Neuroligins

and

neurexins

link synaptic function to cognitive disease. Nature 455, 903–911 (2008).

Iossifov

et al., De novo gene disruptions in children on the autistic spectrum. Neuron, 2012, 74, 285–299

R. Bernier et al., Disruptive CHD8 mutations define a subtype of autism early in development. Cell, 2014, 158, 263–276

J. Castro, N.

Mellios

, M. Sur, Mechanisms and therapeutic challenges in autism spectrum disorders: Insights from

Rett

syndrome.

Curr

.

Opin

. Neurol. 26, 154–159 (2013).

X. Nan, F. J.

Campoy

, A. Bird, MeCP2 is a transcriptional repressor with abundant binding sites in genomic chromatin. Cell 88, 471–481 (1997

R. R. Meehan, J. D. Lewis, A. P. Bird, Characterization of MeCP2, a vertebrate DNA binding protein with affinity for

methylated

DNA. Nucleic Acids Res. 20, 5085–5092 (1992).

References

Grant proposal

Slide26

26

P. L. Jones et al.,

Methylated

DNA and MeCP2 recruit

histone deacetylase to repress transcription. Nat. Genet. 1997, 19, 187–191

M. Chahrour

et al., MeCP2, a key contributor to neurological disease, activates and represses transcription. Science 2008 320, 1224–1229.

S. Ben-

Shachar

, M.

Chahrour

, C.

Thaller

, C. A. Shaw, H. Y.

Zoghbi

, Mouse models of MeCP2 disorders share gene expression changes in the cerebellum and hypothalamus.

Hum.Mol

. Genet. 2009, 18, 2431–2442 .

R. G.

Urdinguio

et al., Disrupted

microRNA

expression caused by Mecp2 loss in a mouse model of

Rett

syndrome.

Epigenetics

2010 5, 656–663 .

H. Wu et al., Genome-wide analysis reveals methyl-

CpGbinding

protein 2-dependent regulation of

microRNAs

in a mouse model of

Rett

syndrome. Proc. Natl. Acad. Sci. U.S.A. 2010, 107, 18161–18166

P. Jin et al., Biochemical and genetic interaction between the fragile X mental retardation protein and the

microRNA

pathway. Nat.

Neurosci

. 2004, 7, 113–117

J. O. Lipton, M.

Sahin

, The neurology of

mTOR

. Neuron2014, 84, 275–291.

M. Qin et al., Altered cerebral protein synthesis in fragile X syndrome: Studies in human subjects and knockout mice. J.

Cereb

. Blood Flow

Metab

. 2014, 33, 499–507 .

References

Grant proposal

Slide27

27

J. C. Darnell et al., FMRP stalls ribosomal translocation on mRNAs linked to synaptic function and autism. Cell 2011, 146, 247–261 .

Q. Chang, G.

Khare

, V.

Dani, S. Nelson, R. Jaenisch, The disease progression of Mecp2 mutant mice is affected by the level of BDNF expression. Neuron 2006, 49, 341–348.

D.

Tropea

et al., Partial reversal of

Rett

Syndrome-like symptoms in MeCP2 mutant mice. Proc. Natl. Acad. Sci. U.S.A. 2009, 106, 2029–2034.

N.

Mellios

et al., b2-Adrenergic receptor agonist ameliorates phenotypes and corrects

microRNA

-mediated IGF1 deficits in a mouse model of

Rett

syndrome. Proc. Natl. Acad. Sci. U.S.A. 2014, 111, 9947–9952

J. Castro et al., Functional recovery with recombinant human IGF1 treatment in a mouse model of

Rett

Syndrome. Proc. Natl. Acad. Sci. U.S.A. 2014, 111, 9941–9946.

S.

Ricciardi

et al., Reduced AKT/

mTOR

signaling and protein synthesis

dysregulation

in a

Rett

syndrome animal model. Hum. Mol. Genet. 2011, 20, 1182–1196.

Kim KC,

Choi

CS, Kim JW, Han SH, Cheong JH,

Ryu

JH, Shin CY, MeCP2 modulates sex differences in the postsynaptic development of the

valproate

animal model of autism. 2011, Mol

Neurobiol

.

Kim KC, Kim P, Go HS,

Choi

CS, Park JH, Kim HJ,

Jeon

SJ,

Dela

Pena IC, Han SH, Cheong JH,

Ryu

JH, Shin CY. Male-specific alteration in excitatory post-synaptic development and social interaction in pre-natal

valproic

acid exposure model of autism spectrum disorder. J

Neurochem

2013, 124:832-843.

References

Grant proposal

Slide28

28

Kim KC, Kim P, Go HS,

Choi

CS, Yang SI, Cheong JH, Shin CY,

Ko KH, The critical period of valproate

exposure to induce autistic symptoms in Sprague-Dawley rats.

Toxicol

Lett

2011, 201:137-142.

Kim KC, Lee DK, Go HS, Kim P,

Choi

CS, Kim JW,

Jeon

SJ, Song MR, Shin CY. Pax6-dependent cortical

glutamatergic

neuronal differentiation regulates autism-like behavior in prenatally

valproic

acid-exposed rat offspring. Mol

Neurobiol

2014, 49:512-528.

Bourin

M, Petit-

Demoulière

B,

Dhonnchadha

BN,

Hascöet

M Animal models of anxiety in mice.

Fundam

Clin

Pharmacol

2007, 21:567-574.

Epstein DH, Preston KL, Stewart J,

Shaham

Y Toward a model of drug relapse: an assessment of the validity of the reinstatement procedure. Psychopharmacology (

Berl

) 2006, 189:1-16.

Lubow

RE. Construct validity of the animal latent inhibition model of selective attention deficits in schizophrenia.

Schizophr

Bull 2005, 31:139-153.

Rodier

PM, Ingram JL, Tisdale B, Nelson S, Romano J, Embryological origin for autism: developmental anomalies of the cranial nerve motor nuclei. J Comp

Neurol

1996, 370:247-261.

References

Grant proposal

Slide29

29

Williams G, King J, Cunningham M, Stephan M, Kerr B,

Hersh

JH Fetal

valproate syndrome and autism: additional evidence of an association. Dev Med Child Neurol2001¸ 43:202-206

Schneider T, Przewłocki

R Behavioral alterations in rats prenatally exposed to

valproic

acid: animal model of autism.

Neuropsychopharmacology

2005, 30:80-89.

Volkmar

FR,

Pauls

D, Autism. Lancet 2003, 362:1133-1141.

Lord C,

Risi

S,

Lambrecht

L, Cook EH

Jr

,

Leventhal

BL,

DiLavore

PC, Pickles A,

Rutter

M. The autism diagnostic observation schedule-generic: a standard measure of social and communication deficits associated with the spectrum of autism. J Autism Dev

Disord

2000, 30:205-223.

Wing L, Gould J, Severe impairments of social interaction and associated abnormalities in children: epidemiology and classification. J Autism Dev

Disord

1979, 9:11-29.

Belzung

C,

Lemoine

M (2011) Criteria of validity for animal models of psychiatric disorders: focus on anxiety disorders and depression.

Biol

Mood Anxiety

Disord

1:9.

Dawson G,

Meltzoff

AN,

Osterling

J,

Rinaldi

J, Brown E, Children with autism fail to orient to naturally occurring social stimuli. J Autism Dev

Disord

1998, 28:479-485.

Kasari

C, Patterson S, Interventions addressing social impairment in autism.

Curr

Psychiatry Rep 2012, 14:713-725.

Ganaie

SA,

Bashir

A.

Global Autism: Autism, Autism Etiology, Perceptions, Epistemology, Prevalence and Action.

Int

J

Clin

Ther

Diagn

. 2(2), 39-47.

References

Grant proposal