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Estrogen reduces TLR signaling �in monocytes

Tanya Freedman Lab

Wednesday, August 16th

Her, Zuag Paj

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Some autoimmune diseases have a female-bias

Female Bias

Autoimmune Disease

Infectious Disease

Non-reproductive cancers

  • Systemic Lupus Erythematosus
  • Rheumatoid Arthritis
  • Multiple Sclerosis
  • Type 1 Diabetes
  • Graves Disease
  • HIV
  • Influenza
  • Malaria
  • Zika
  • Toxoplasmosis
  • Legionella
  • Bladder
  • Bowel
  • Kidney
  • Leukemia
  • Liver
  • Lung

Sauer, M.

Adapted from Klein and Flanagan., Nature Rev. Imm., 2016

​

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Some autoimmune diseases have a female-bias

Female Bias

Autoimmune Disease

Infectious Disease

Non-reproductive cancers

  • Systemic Lupus Erythematosus
  • Rheumatoid Arthritis
  • Multiple Sclerosis
  • Type 1 Diabetes
  • Graves Disease
  • HIV
  • Influenza
  • Malaria
  • Zika
  • Toxoplasmosis
  • Legionella
  • Bladder
  • Bowel
  • Kidney
  • Leukemia
  • Liver
  • Lung

Sauer, M.

Adapted from Klein and Flanagan., Nature Rev. Imm., 2016

​

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Systemic Lupus Erythematosus (SLE)

Adapted from Jiang, W.

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Loss of Src-family kinase Lyn drives lupus

LynAKO

LynKO

WT

​

Brian, Sauer,…Freedman., Sci. Adv., 2022

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Female LynAKO mice have increased numbers of pro-inflammatory macrophages and severe splenomegaly

​

Brian, Sauer,…Freedman., Sci. Adv., 2022

Average fluorescence of flow cytometry (rel. to WT)

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Lyn regulation of Toll-like receptors (TLRs) remains unclear

Adapted from Brian and Freedman, Endocrinology., 2021

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Adapted from Brian and Freedman, Endocrinology., 2021

Lyn regulation of Toll-like receptors (TLRs) remains unclear

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Lyn regulation of Toll-like receptors (TLRs) remains unclear

Adapted from Brian and Freedman, Endocrinology., 2021

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Could estrogen induce TLR expression and mask the inhibitory regulation of Lyn?

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Estrogen Receptor (ER) signaling

Adapted from Słowikowski et al., Mol Biol Rep., 2017

17β-Estradiol (E2)

ER

Src

P

P

MNAR

ER

ER

∆ transcription

and proliferation

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Estrogen may increase inflammation through upregulation of TLR signaling

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Adapted from Sauer, M.

Estrogen may increase inflammation through upregulation of TLR

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Adapted from Sauer, M.

Estrogen may increase inflammation through upregulation of TLR

LynAKO

LynAKO

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How will we look at the effect of estrogen on TLR signaling in monocytes?

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Experimental workflow of TLR agonist stimulations

pIKK⍺/β and pAKT

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NS

Timepoint (min):

0

30

45

60

75

90

NS

0

Na3VO4

pIKK⍺/β

(S176/180)

b-Actin

5

5

30

45

60

75

90

E2

(+) LPS, 100 ng/mL

pIKK⍺/β / TPS rel. to t=0

N=5

E2, 10 nM

untreated

Estrogen (E2) reduces pIKK⍺/β and pAKT levels in TLR4 signaling

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NS

Timepoint (min):

0

30

45

60

75

90

NS

0

Na3VO4

pIKK⍺/β

(S176/180)

pAKT

(s473)

b-Actin

5

5

30

45

60

75

90

E2

(+) LPS, 100 ng/mL

pIKK⍺/β / TPS rel. to t=0

pAKT / TPS rel. to t=0

N=5

Estrogen (E2) reduces pIKK⍺/β and pAKT levels in TLR4 signaling

E2, 10 nM

untreated

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Estrogen reduces activation of IKK⍺/β and AKT

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Next Steps

LynAKO

LynKO

WT

Next Steps

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Thank you & Questions

Figures were made with

Freedman Lab

PI: Tanya Freedman

Monica Sauer

J.T. Greene

Will Kanagy

Anders Lindstedt

Erandika Senevirathne

Silvia Toledo

Catherine Duffy Shaw

Medical Student Summer Research Program in Infection and Immunity

Director: Daniel Mueller

Administrator: Drew Keup

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Estrogen reduces TLR4 signaling in monocytes