13alzforum carson

8
Monica J Carson ([email protected] ) Microglia as a CNSspecific 9ssue macrophage: TREM2 dependent func9ons TREM2 DAP12/ TRYROBP Tmem176b C1qA ! "#$%&'()*+& ,-./01-/. 23%41$5 "# $ %&'() *"+,) -- .%"/ 123456243 78(524

Upload: alzheimer-research-forum

Post on 07-Jul-2015

3.122 views

Category:

Technology


1 download

TRANSCRIPT

Page 1: 13alzforum carson

Monica  J  Carson  ([email protected])  

Microglia  as  a  CNS-­‐specific  9ssue  macrophage:    TREM2  dependent  func9ons  

TREM2

DAP12/ TRYROBP

Tmem176b

C1qA

!

!!!!"#$%&'()*+&!

,-./01-/.!23%41$5

"# $#

%&'()

*"+,)

--.%"/0123456243078(524

%&'()

%('(,9:;

< < <= = =!0000>000!0000>0000!0000>000!0000>0000!0000>0000!000>

?0(84@A B0(84@A ,C0(84@A!#

!"

#"

$"

%"

&"

'!"

'#"

'$"

'%"

'&"

(")*+,-." /")*+,-." '0")*+,-."

!"

'!"

#!"

(!"

$!"

0!"

%!"

1!"

&!"

/!"

'!!"

(")*+,-." /")*+,-." '0")*+,-."

*# Trem2

Tmem176bD#

Blue=non-transgenic wild-type; Red=APP23 transgenic

Page 2: 13alzforum carson

Microglia are a distinct CNS-specific myeloid lineage but the CNS is also served by infiltrating myeloid cells

B. IC LPS injection

CD45 à

MG MP

A. Untreated, no inflammation

CD45 à

FcR

à

MG

FcR

à

1)  CNS-resident Microglia Microglia are: ü  CD45lo to CD45intermediate ü  long-lived, ü  largely self-replenishing ü  PU.1/IRF8 dependent lineage that

begin populating the CNS during embryonic development

ü  Do not traffic to draining lymph nodes (at detectable rates)

2)  Peripheral monocytes /macrophages

that acutely infiltrate the CNS These cells are: ü  CD45hi ü  short lived ü  rapidly replaced by bone marrow

derived cells in the adult ü  Can traffic to draining lymph nodes

Page 3: 13alzforum carson

Trem1  

Trem2   Macrophage  

Trem2  Trem1  

Microglia  

Bkgrd  

Bkgrd  

mg

100 101 102 103 104

FL2-H

MJC050212.039 11.8MJC050212.030 11.4MJC050212.021 15.7MJC050212.012 11.2

live no boxesMJC050212.011Event Count: 27279

100 101 102 103 104

FL4-H

100

101

102

103

104

FL1-H

CD45 à

FcR

à

C. IP LPS injection MP

MG

Page 4: 13alzforum carson

So what do we know about TREM2 and neuroinflammation:

TREM2 up regulated

Cuprizone

Facial Axotomy In general, situations with increased cell

damage, changes in neuronal activity and/or TNF lead to increased microglial expression of

TREM2

Cell debris or TNF alone each are sufficient to increase microglial expression of TREM2

(Schmid et al. JNC 2002, 2009) Melchior et al. ASN NEURO 2010)

Please note the gene:

DAP12 = TYROBP = KARAP

!

!!!!"#$%&'()*+&!

,-./01-/.!23%41$5

"# $#

%&'()

*"+,)

--.%"/0123456243078(524

%&'()

%('(,9:;

< < <= = =!0000>000!0000>0000!0000>000!0000>0000!0000>0000!000>

?0(84@A B0(84@A ,C0(84@A!#

!"

#"

$"

%"

&"

'!"

'#"

'$"

'%"

'&"

(")*+,-." /")*+,-." '0")*+,-."

!"

'!"

#!"

(!"

$!"

0!"

%!"

1!"

&!"

/!"

'!!"

(")*+,-." /")*+,-." '0")*+,-."

*# Trem2

Tmem176bD#

Blue=non-transgenic wild-type; Red=APP23 transgenic

Page 5: 13alzforum carson

: Why focus on TREM2 in AD Associated Neuroinflammation?

Because  Trem2  is  a  receptor  that:    1)  10-­‐50  fold  higher  on  microglia  than  on  blood-­‐derived  macrophages  2)  In  the  murine  CNS  is  only  expressed  by  microglia  (no  TREM2  in  PU.1  KO’s  lacking  

microglia)    3)  Inhibits  signaling  of  some  pro-­‐inflammatory  receptors  (TLR4)  4)  Increases  phagocytosis  of  cell  debris  and  amyloid  by  microglia        5)  promotes  9ssue  repair  following  injury  and    inflammatory  insults  (Overexpression  &  

an9body  blocking  studies)  6)  TREM2  binding  ac9vity  (puta9ve  TREM2  ligands)  expressed  by  microglia,  CNS-­‐

infiltra9ng  macrophages,  ac9vated  glia  and  stressed  &  apopto9c  neurons  and  glia  

Loss of functional TREM2 signaling leads to early onset cognitive dementia

(20’s) & death by the 40-50’s: Nasu-Hakola Disease

Tempting to speculate that normal age-

related decrease in TREM2 levels contributes to age-related susceptibility

for AD (Thrash et al. 2009)

!"" !"! !"# !"$ !"%

!"#$%&'(&)(&*+),&)'-,.*/0$1*2(3"&4

B: TREM2

PD2

3 mo.

PD21

PD7

Bkgrd

5*$6*738

!"" !"! !"# !"$ !"%

5*$6*738

PD2

PD21

PD7

Bkgrd

C: Mannose Receptor

!"#$%&'(&)(&*+),&)'-,.*/0$1*2(3"&4!"" !"! !"# !"$ !"%

5*$6*738

!"#$%&'(&)(&*+),&)'-,.*/0$1*2(3"&4

PD2

3 mo.

PD21

PD7

Bkgrd

D: PD-L1

!"" !"! !"# !"$ !"%

5*$6*738

!"#$%&'(&)(&*+),&)'-,.*/0$1*2(3"&4

F: MHC Class II

PD2

PD21

PD7

Bkgrd

!"" !"! !"# !"$ !"%

5*$6*738

!"#$%&'(&)(&*+),&)'-,.*/0$1*2(3"&4

G: CD40

PD2

PD21

PD7

Bkgrd!"" !"! !"# !"$ !"%

5*$6*738

!"#$%&'(&)(&*+),&)'-,.*/0$1*2(3"&4

H. B7.2

PD2

PD21

PD7

Bkgrd

!"" !"! !"# !"$ !"%

5*$6*738

!"#$%&'(&)(&*+),&)'-,.*/0$1*2(3"&4

E: CX3CR1

PD3

PD21

PD14

Bkgrd

3 mo.

!"" !"! !"# !"$ !"%!""

!"!

!"#

!"$

!"%

!&'(

)*+,-./*0

9:;<

9:==>

A.

Page 6: 13alzforum carson

Microglia are the tissue macrophage of the brain…. So, what are the roles and functions of the typical tissue macrophage?

Is the recent AD-associated mutation in TREM2 ligand binding domain, loss of function or altered recognition of ligands leading to dysregulated TREM2 dependent microglial activation?

Phagocytosis

Antigen-presentation To T cells: activate, retain direct effector functions

Trafficking to lymph nodes

Tissue homeostasis

Tissue repair

Pathogen defense

Microglia not detected to traffick to cervical lymph

nodes

Microglia: yes TREM2 regulated

Microglia: Yes: TREM2 regulated

Microglia: Yes via innate and adaptive

immunity TREM2 regulated

Microglia: Yes, But as inhibitory/tolerogenic APCs

TREM2 regulated in vitro

Microglia: Yes, very active innate immunity and oxidative pathways

TREM2 regulated

Page 7: 13alzforum carson

TREM2 immunoreactivity: a caution

!

!!!!"#$%&'()*+&!

,-./01-/.!23%41$5

"# $#

%&'()

*"+,)

--.%"/0123456243078(524

%&'()

%('(,9:;

< < <= = =!0000>000!0000>0000!0000>000!0000>0000!0000>0000!000>

?0(84@A B0(84@A ,C0(84@A!#

!"

#"

$"

%"

&"

'!"

'#"

'$"

'%"

'&"

(")*+,-." /")*+,-." '0")*+,-."

!"

'!"

#!"

(!"

$!"

0!"

%!"

1!"

&!"

/!"

'!!"

(")*+,-." /")*+,-." '0")*+,-."

*# Trem2

Tmem176bD#

Blue=non-transgenic wild-type; Red=APP23 transgenic

Microglia also produce soluble “decoy” form of receptor! Therefore reports of TREM2 immunoreactivity on non-microglia might reflect detection of the soluble variant of TREM2 binding these TREM2 ligands on neurons, glia and

perivascular macrophages Unpublished results, Schmid et al. 2002, Hamerman et al 2006,

Piccio et al. 2008, Hsieh et al.2009

TREM2 binding activity (putative ligand expression) is detected on reactive astrocytes, infiltrating macrophages (high levels) and stressed and/or apoptotic neurons, reactive and apoptotic glia

Page 8: 13alzforum carson

NINDS UCR Div of Biomed Sci PIC grant

UCR Chancellor’s Strategic Initiative Dana Foundation

Biogen Idec, Merck DOD; CRCC

Tammy Kielian

Caroline Whitacre

Emma Wilson

Kathryn Jones Virginia Sanders

Marco Colonna

Harald Neuman

Jenny Ting

Carson Lab Tiffany Butts

Deirdre S. Davis Alfredo Hernandez

Benoit Melchior Yoshinori Otani

Shweta Puntambekar Victoria Senechal

Christoph D. Schmid J. Cameron Thrash

Iryna M. Ethell Tina Bilousova Slawomir Sloniwski Peter Hickmott Devin Binder Mike Hsu

Microglia and Macrophages in brain development and neuroinflammation