Ja FSH Receptor

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Presentation Description

Presented by William Ja at the Alliance for Contraception in Cats & Dogs’ 4th International Symposium on Non-Surgical Contraceptive Methods of Pet Population Control, April 8-10, 2010, in Dallas, Texas, U.S.

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Presentation Transcript

Slide 1: 

William W. Ja The Scripps Research Institute Jupiter, Florida FSH receptor ligand-cytotoxin conjugates: Potential for permanent chemosterilization

The Scripps Research Institute : 

The Scripps Research Institute

Found Animals Foundation : 

Found Animals Foundation

Found Animals Foundation : 

Found Animals Foundation

Targeting the reproductive system : 

Targeting the reproductive system Grimm, 2009

Targeting the reproductive system : 

Targeting the reproductive system Grimm, 2009 Emanuele, 2003

Targeted cytotoxins : 

Targeted cytotoxins Targeting ligand Cytotoxin

Targeted cytotoxins : 

Targeted cytotoxins Targeting ligand Cytotoxin High potency High specificity No off-target effects!!

Can we target gonadal cells critical to reproduction? : 

Can we target gonadal cells critical to reproduction?

Can we target gonadal cells critical to reproduction? : 

Can we target gonadal cells critical to reproduction?

Can we target gonadal cells critical to reproduction? : 

Can we target gonadal cells critical to reproduction? gonadotropins

Target: Follicle-stimulating hormone receptor : 

Target: Follicle-stimulating hormone receptor Wang, 2001 Gapd FSHR

Timeline : 

Timeline 1950 1960 1970 1980 1990 2000 Gonadotropins used to treat ovulatory dysfunction

Timeline : 

Timeline 1950 1960 1970 1980 1990 2000 Gonadotropins used to treat ovulatory dysfunction Binding of FSH to gonadal sites

Timeline : 

Timeline 1950 1960 1970 1980 1990 2000 Gonadotropins used to treat ovulatory dysfunction Binding of FSH to gonadal sites Classical biochemistry

Timeline : 

Timeline 1950 1960 1970 1980 1990 2000 Gonadotropins used to treat ovulatory dysfunction Binding of FSH to gonadal sites FSHR cDNA cloned Classical biochemistry

Timeline : 

Timeline 1950 1960 1970 1980 1990 2000 Gonadotropins used to treat ovulatory dysfunction Binding of FSH to gonadal sites FSHR cDNA cloned FSH-FSHR structure Classical biochemistry

FSHR: Sertoli cells : 

FSHR: Sertoli cells Gray’s Anatomy

FSHR: Sertoli cells : 

FSHR: Sertoli cells Gray’s Anatomy

FSHR: granulosa cells : 

FSHR: granulosa cells Gray’s Anatomy

Sertoli & granulosa cells are crucial for reproduction : 

Sertoli & granulosa cells are crucial for reproduction

Sertoli & granulosa cells are crucial for reproduction : 

Sertoli & granulosa cells are crucial for reproduction

G protein-coupled receptors : 

G protein-coupled receptors Klaasse, 2008

G protein-coupled receptors : 

G protein-coupled receptors Klaasse, 2008 Accessible Drug-able Internalize

FSH is a heterodimer : 

FSH is a heterodimer α FSHb

FSHb is amenable to C-terminal conjugation : 

FSHb is amenable to C-terminal conjugation (PDB ID: 1XWD (Fan, 2005) FSHR ectodomain FSHa FSHb

Stable mammalian cell lines : 

Stable mammalian cell lines Selection on hygromycin B & geneticin

FSH expression : 

FSH expression media cells Secreted Non-secreted

FSH expression : 

FSH expression 25 kDa media cells cells media FSHβ FSHα 20 kDa

Choosing a cytotoxin : 

Choosing a cytotoxin Exotoxin A Diphtheria toxin Ricin Shiga toxin Cell-binding Cell-killing

Choosing a cytotoxin : 

Choosing a cytotoxin Exotoxin A Diphtheria toxin Ricin Shiga toxin Cell-binding Cell-killing High potency Required internalization Survive proteolytic processing Economically produced (E. coli)

Choosing a cytotoxin : 

Choosing a cytotoxin Cell-killing Target-binding Exotoxin A Diphtheria toxin Ricin Shiga toxin High potency Required internalization Survive proteolytic processing Economically produced (E. coli)

Exotoxin A : 

Exotoxin A Cell-binding Cell-killing Translocation ADP-ribosylation I II III II III PE38

Exotoxin A entry : 

Exotoxin A entry Pastan, 2006 ExoA cell-killing Targeting ligand

Exotoxin A : 

Exotoxin A Cell-binding Translocation I II III II III PE38 ADP-ribosylation

Exotoxin A : 

Exotoxin A Cell-binding Translocation I II III II III PE38 Target-binding ADP-ribosylation

Peptide “velcro” for conjugation : 

Peptide “velcro” for conjugation basic peptide acidic peptide O’Shea, 1993

Peptide “velcro” for conjugation : 

Peptide “velcro” for conjugation basic peptide acidic peptide O’Shea, 1993 SH HS

Peptide “velcro” for conjugation : 

Peptide “velcro” for conjugation basic peptide acidic peptide O’Shea, 1993 S S

PE38 for conjugation : 

PE38 PE38 for conjugation

FSH-PE38 : 

II III α FSHb PE38 FSH-PE38

FSHR-FSH structure : 

FSHR-FSH structure (PDB ID: 1XWD (Fan, 2005) FSHR ectodomain FSHa FSHb

FSHb peptide mimetics : 

FSHb peptide mimetics (PDB ID: 1XWD (Fan, 2005)

FSHb peptide mimetics : 

FSHb peptide mimetics

FSHb peptide mimetics : 

FSHb peptide mimetics

Modular cytotoxin conjugates : 

Modular cytotoxin conjugates

Modular cytotoxin conjugates : 

Modular cytotoxin conjugates

Off-target effects: urinary tract : 

Off-target effects: urinary tract Control FSHR Ponglowhapan, 2007

Off-target effects: VLS : 

Off-target effects: VLS II III PE38

Off-target effects: VLS : 

Off-target effects: VLS II III PE38 Non-specific toxicity Wang, 2007

Off-target effects: VLS : 

Off-target effects: VLS II III PE38 Non-specific toxicity Proteolysis Immunogenicity Wang, 2007

Exotoxin A : 

Exotoxin A II III PE38 Wang, 2007 pDW363C Non-specific toxicity Proteolysis Immunogenicity

Timeline : 

Timeline FSH PE38

Acknowledgments : 

Acknowledgments Megan Larisch (Caltech) Claudia Harden (Palm Beach Zoo) David Waugh (NCI) Found Animals Foundation wja@scripps.edu

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