CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara...

55
CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University

TAGS:

Transcript of CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara...

Page 1: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN

HEAD AND NECK CANCER

Devathri NanayakkaraEskitis Institute for Drug Discovery

Griffith University

Page 2: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Head and neck cancer

Page 3: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Head and neck cancer

Sixth most common cancer

Page 4: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Head and neck cancer

Sixth most common cancer

Five year survival rate after diagnosis, 50%

Page 5: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Head and neck cancer

Sixth most common cancer

Five year survival rate after diagnosis, 50%

Drug resistance

Early tumors - asymptomatic

Page 6: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Head and neck cancer

Sixth most common cancer

Five year survival rate after diagnosis, 50%

Drug resistance

Early tumors - asymptomatic

“Need to study the underlying molecular pathways unveiling potential detection markers and drug targets”

Page 7: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

In a recent study,

• Characterized the somatic mutation landscape of OSCC-GB

Page 8: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.
Page 9: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

- Found Five new genes associated with OSCC-GB

Page 10: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

- Among them was USP9X- 22% harboured copy number loss and truncating

mutations - Role as tumor suppressor

Page 11: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

USP9X• Deubiquitylating enzyme– Family of cysteine and metalloproteases

Page 12: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

USP9X• Deubiquitylating enzyme– Family of cysteine and metalloproteases

http://legacy.butler.edu/biology/faculty-staff/research-interests/jkowalski/research-in-the-kowalski-lab/

Page 13: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Murtaza et al, 2015

Page 14: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

USP9X in cancer

Page 15: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

USP9X in cancer

Page 16: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

USP9X in cancerBreast cancer

Colorectal cancer

Bladder cancer

Oral cancer

Brain cancer

Pancreatic cancer

Prostate cancer

Lung cancer

Lymphoma

Page 17: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

In this study,

Aims :• To evaluate the role of USP9X in an in vitro

system• To elucidate the molecular mechanisms USP9X

is involved in

Page 18: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

How?• In vitro cell lines – SCC15, CAL27, FaDu and Detroit 562

Page 19: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Immunoblotting to probe for USP9X expression

All 4 cell lines express USP9X

SCC1

5

FaD

u

Det

roit

562

CAL2

7

USP9X290 kDa

β tubulin51 kDa

Page 20: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

• Knockdown approach – siRNA

Page 21: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Immunoblotting to probe for USP9X protein levels 72 h after siRNA treatment

USP9X is efficiently knocked down in all four cell lines

NT USP9X NT USP9X NT USP9X NT USP9X

SCC15 CAL27 FaDu Detroit 562

siRNA

USP9X290 kDa

Β tubulin51 kDa

Page 22: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

• Effect on cell aspects– Cell proliferation• CyQUANT Assay

Page 23: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

48H 72H 96H 120H 144H0

2

4

6

8

10

12

14

16

18

NT siRNA

USP9x siRNA

Nor

mal

ised

fluor

esce

nce

48H 72H 96H 120H 144H0

5

10

15

20

25

30

NT siRNAUSP9X siRNA

Nor

mal

ised

fluor

esce

nce

48H 72H 96H 120H 144H0

2

4

6

8

10

12

14

16

NT siRNAUSP9X siRNA

Nor

mal

ised

fluor

esce

nce

48H 72H 96H 120H 144H0

2

4

6

8

10

12

14

NT siRNAUSP9X siRNA

Nor

mal

ised

fluor

esen

ce

time time

time time

In absence of USP9X, a decrease in cell numbers was observed

CyQUANT Analysis of cell proliferation following siRNA treatment to knockdown USP9X

SCC15

Detroit 562

CAL27

FaDu

*

*

* * *

*

*

*

* P value < 0.05

Page 24: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

• Decrease in cell numbers,– Apoptosis?

Page 25: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

No elevation in apoptosis detected upon depletion of USP9X

β tubulin 51 kDa

Cleaved PARP-1 89 kDa

siRNA NT USP9X NT USP9X NT USP9X NT USP9X NT USP9X NT USP9X NT USP9X NT USP9X NT USP9X NT USP9X NT USP9X NT USP9X

time 48h 96h 144h 48h 96h 144h 48h 96h 144h 48h 96h 144h

SCC15 CAL27 FaDu Detroit 562

Immunoblotting for cleaved PARP-1

Page 26: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Decrease in cell numbers in absence of USP9X prompts a role of a

Page 27: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Decrease in cell numbers in absence of USP9X prompts a role of a “tumor promoter”

Page 28: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Decrease in cell numbers in absence of USP9X prompts a role of a “tumor promoter”• Contradicts predicted oncosupressive role• Context specific

Page 29: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Decrease in cell numbers in absence of USP9X prompts a role of a “tumor promoter”• Contradicts predicted oncosupressive role• Context specific

– Pancreatic cancer

Page 30: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Decrease in cell numbers in absence of USP9X prompts a role of a “tumor promoter”• Contradicts predicted oncosupressive role• Context specific

– Pancreatic cancer

Page 31: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Decrease in cell numbers in absence of USP9X prompts a role of a “tumor promoter”• Contradicts predicted oncosupressive role• Context specific

– Pancreatic cancer

Tumor suppressor

Page 32: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Decrease in cell numbers in absence of USP9X prompts a role of a “tumor promoter”• Contradicts predicted oncosupressive role• Context specific

– Pancreatic cancer

Tumor suppressor

Page 33: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Decrease in cell numbers in absence of USP9X prompts a role of a “tumor promoter”• Contradicts predicted oncosupressive role• Context specific

– Pancreatic cancer

Tumor suppressor

Tumor promotor

Page 34: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

To further confirm,• Overexpression of USP9X

Page 35: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

To further confirm,• Overexpression of USP9X

Usp9x cDNA

Page 36: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

To further confirm,• Overexpression of USP9X

Usp9x cDNA

Linearized the plasmid

Lipofectamine 2000

After 3 days: antibiotic selection(12 days)

To establish stable cell lines,

Page 37: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

USP9X is ectopically expressed in all four cell lines

V5SC

C15

CAL2

7

FaDu

Detr

oit 5

62

scc1

5

CAL2

7

FaDu

Detr

oit 5

62

pDEST51 USP9X pDEST51

β tubulin

Immunoblotting to detect ectopic expression of USP9X

Page 38: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

48h 72h 96h 120h 144h0

2

4

6

8

10

12

14

16

pdest51pdest51 USP9X

Nor

mal

ised

fluor

esen

ce

48h 72h 96h 120h 144h0

5

10

15

20

25

30

pdest51pdest51 USP9X

Nor

mal

ised

fluor

esen

ce

48h 72h 96h 120h 144h0

2

4

6

8

10

12

14

pdest51pdest51 USP9X

Nor

mal

ised

fluor

esen

ce

48h 72h 96h 120h 144h02468

101214161820

pdest51pdest51 USP9X

Nor

mal

ised

fluor

esen

ce

Ectopic USP9X protein expression increased cell proliferation

CyQUANT Analysis of cell proliferation following ectopic expression of USP9X

SCC15

Detroit 562

CAL27

FaDu

*

* *

*

* *

**

*

*

* *

*

* P value < 0.05

Page 39: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Cell numbers are directly proportional to level of USP9X protein

Page 40: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Cell numbers are directly proportional to level of USP9X protein has an oncogenic role

Page 41: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Molecular mechanism regulated by USP9X

Page 42: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Molecular mechanism regulated by USP9X

Murtaza et al, 2015

Page 43: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Molecular mechanism regulated by USP9X– mTOR– Wnt – Notch

• Regulates cell proliferation• Known USP9X substrates

Page 44: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Molecular mechanism regulated by USP9X?– mTOR– Wnt – Notch

CyclinD1, c-MYC, HES1

Page 45: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Molecular mechanism regulated by USP9X?– mTOR– Wnt – Notch

Quantitate the RNA levels by qPCR

CyclinD1, c-MYC, HES1

Page 46: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

48H 72H 96H 120H 144H0

2

4

6

8

10

12

14

16

18

NT siRNA

USP9x siRNA

Nor

mal

ised

fluor

esce

nce

48h 72h 96h 120h 144h0

2

4

6

8

10

12

14

16

pdest51pdest51 USP9X

Nor

mal

ised

fluor

esen

ce

RNA extraction

cDNA

qPCR

Page 47: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Fold change of target genes 144 h after knockdown of USP9X

SCC15 CAL27

FaDu Detroit 562

Page 48: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Fold change of target genes 144 h after knockdown of USP9X

SCC15 CAL27

FaDu Detroit 562

Page 49: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Fold change of target genes 144 h after knockdown of USP9X

SCC15 CAL27

FaDu Detroit 562

48H 72H 96H 120H 144H0

5

10

15

20

25

30

NT siRNAUSP9X siRNA

Nor

mal

ised

fluor

esce

nce

Page 50: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

SCC15 CAL27

FaDu Detroit 562

Fold change of target genes 144 h after ectopic expression of USP9X

Page 51: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

• Consistently HES1 expression correlated with cell proliferation measured by CyQUANT assay

• USP9X seems to positively regulate notch pathway

Page 52: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

• Consistently HES1 expression correlated with cell proliferation measured by CyQUANT assay

• USP9X seems to positively regulate notch pathway

Hypothesis: USP9X regulates proliferation of head and neck cancer cells through notch pathway

Page 53: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

Conclusions• USP9X depletion caused a decrease in cell

proliferation• Ectopic expression of USP9X led to increase in

cell proliferation• USP9X positively regulates Notch pathway

Page 54: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

ACKNOWLEDGEMENT

StephenGeorge

Nicholas SaundersWood lab members

Funding: Griffith University

Page 55: CONTEXT SPECIFIC ROLE OF DEUBIQUITYLASE ENZYME, USP9X, IN HEAD AND NECK CANCER Devathri Nanayakkara Eskitis Institute for Drug Discovery Griffith University.

THANK YOU!


FAM/USP9x, a Deubiquitinating Enzyme …snewfeld.lab.asu.edu/pdf/newfeld_fam_print.pdfFAM/USP9x, a Deubiquitinating Enzyme Essential for TGFb Signaling, Controls Smad4 Monoubiquitination

FAM/USP9x, a Deubiquitinating Enzyme …snewfeld.lab.asu.edu/pdf/newfeld_fam_print.pdfFAM/USP9x, a Deubiquitinating Enzyme Essential for TGFb Signaling, Controls Smad4 Monoubiquitination

Intellectual Property Issues in Exporting Tamara Nanayakkara Counsellor SMEs Division.

Intellectual Property Issues in Exporting Tamara Nanayakkara Counsellor SMEs Division.

LICENSING AND FRANCHISING; FUNDAMENTALS Tamara Nanayakkara.

LICENSING AND FRANCHISING; FUNDAMENTALS Tamara Nanayakkara.

Primitives for Motor Adaptation Reflect Correlated Neural ......Gary C. Sing1, Wilsaan M. Joiner2, Thrishantha Nanayakkara 1, Jordan B. Brayanov & Maurice A. Smith ... perturbation,

Primitives for Motor Adaptation Reflect Correlated Neural ......Gary C. Sing1, Wilsaan M. Joiner2, Thrishantha Nanayakkara 1, Jordan B. Brayanov & Maurice A. Smith ... perturbation,

Negotiating Technology License Agreements Tamara Nanayakkara.

Negotiating Technology License Agreements Tamara Nanayakkara.

THURSDAY, AUGUST 13, 2020 TOTAL CASES COVID-19 TOTAL … · Vasudeva Nanayakkara – Water Supply Udaya Gammanpila – Energy Dr. Ramesh Pathirana – Plantations Prasanna Ranatunga

THURSDAY, AUGUST 13, 2020 TOTAL CASES COVID-19 TOTAL … · Vasudeva Nanayakkara – Water Supply Udaya Gammanpila – Energy Dr. Ramesh Pathirana – Plantations Prasanna Ranatunga

Breaking the chains: deubiquitylase specificity meets ...

Breaking the chains: deubiquitylase specificity meets ...

FAM/USP9x, a Deubiquitinating Enzyme Essential for TGFβ Signaling, Controls Smad4 Monoubiquitination Sirio Dupont, Anant Mamidi, Michelangelo Cordenonsi,

FAM/USP9x, a Deubiquitinating Enzyme Essential for TGFβ Signaling, Controls Smad4 Monoubiquitination Sirio Dupont, Anant Mamidi, Michelangelo Cordenonsi,

Alcohol Prevention by Shakya Nanayakkara

Alcohol Prevention by Shakya Nanayakkara

PROJECT CONCEPT PAPER TEMPLATE - Asian · PDF fileS. Muthugala, Senior Procurement Officer, Sri Lanka Resident Mission, SARD . A. Nanayakkara, Senior Project Officer (Road and Transport),

PROJECT CONCEPT PAPER TEMPLATE - Asian · PDF fileS. Muthugala, Senior Procurement Officer, Sri Lanka Resident Mission, SARD . A. Nanayakkara, Senior Project Officer (Road and Transport),

The Ubiquitin Ligase Itch is Auto-ubiquitylated In Vivo and In Vitro ... · 12-10-2006  · Degradation By Interacting With The Deubiquitylating Enzyme FAM/USP9X*. ... 20 mM NaH 2PO

The Ubiquitin Ligase Itch is Auto-ubiquitylated In Vivo and In Vitro ... · 12-10-2006  · Degradation By Interacting With The Deubiquitylating Enzyme FAM/USP9X*. ... 20 mM NaH 2PO

The Efficacy of Interaction Behavior and Internal Stiffness ...€¦ · Embodied Information Gain in Haptic Perception Nantachai Sornkarn and Thrishantha Nanayakkara Abstract—Haptic

The Efficacy of Interaction Behavior and Internal Stiffness ...€¦ · Embodied Information Gain in Haptic Perception Nantachai Sornkarn and Thrishantha Nanayakkara Abstract—Haptic

Leveraging Intellectual Property Assets for Business Success Tamara Nanayakkara Counsellor Small and Medium Sized Enterprises Division World Intellectual.

Leveraging Intellectual Property Assets for Business Success Tamara Nanayakkara Counsellor Small and Medium Sized Enterprises Division World Intellectual.

Casting Defects- Dr. Manjula Nanayakkara

Casting Defects- Dr. Manjula Nanayakkara

Intellectual Property Issues in Exporting Tamara Nanayakkara Counsellor Small and Medium sized Enterprises Division World Intellectual Property Organization.

Intellectual Property Issues in Exporting Tamara Nanayakkara Counsellor Small and Medium sized Enterprises Division World Intellectual Property Organization.

Supplemental Materials and Methods Supplemental … cells were treated or co-treated with MG132, cycloheximide, or DMSO; or were transfected with siRNAs against USP9X or a non- targeting

Supplemental Materials and Methods Supplemental … cells were treated or co-treated with MG132, cycloheximide, or DMSO; or were transfected with siRNAs against USP9X or a non- targeting

Plant Poisoning Dr. Ravi Nanayakkara. Objectives Introduction, classification and identification of poisonous plants. Circumstances, route of entry, metabolism.

Plant Poisoning Dr. Ravi Nanayakkara. Objectives Introduction, classification and identification of poisonous plants. Circumstances, route of entry, metabolism.

A generic approach for developing dedicated database ... · PDF fileA generic approach for developing dedicated database systems for professional applications L.D.J.F. Nanayakkara

A generic approach for developing dedicated database ... · PDF fileA generic approach for developing dedicated database systems for professional applications L.D.J.F. Nanayakkara

Direct Navigation - Nora Nanayakkara - Sedo

Direct Navigation - Nora Nanayakkara - Sedo

Governmental Influence on Trade Chapter seven Members K.A.S. NavarathnePGIA 07-6906 W.A.Wasantha PGIA 07-6970 R.G.M.S.Perera PGIA 07-6931 J.R. Nanayakkara.

Governmental Influence on Trade Chapter seven Members K.A.S. NavarathnePGIA 07-6906 W.A.Wasantha PGIA 07-6970 R.G.M.S.Perera PGIA 07-6931 J.R. Nanayakkara.