Arrowhead Pharmaceuticals : ATS 2025 RAGE Preclinical 2

Published: 2025-05-18 20:06:54 ET ARWR

Published on 05/18/2025 at 20:06

Timothy N. Perkins, PhD1*, David I Kasahara, PhD2, Taylor Reed2, Holly Hamilton, PhD2, James Hamilton, MD2, MD, Tim D. Oury, MD, PhD1, Erik W. Bush, PhD2

1Deptartment of Pathology, University of Pittsburgh, School of Medicine. Pittsburgh, PA, USA. 2Arrowhead Pharmaceuticals, Madison, WI, USA. *Corresponding author.

RESULTS

Background

RESULTS

Results

Summary

Discussion

Our previous studies have demonstrated that full genetic knockout of RAGE (global) significantly protects mice in several experimental models of asthma

Here we demonstrate that haplodeficiency also partially protects mice several models of T2hi asthma

We have also recently shown that RAGE-deficient mice are protected in a model of severe T2lo asthma. However, studies are needed in additional models to assess the effects of haplodeficiency

Current data suggests that RAGE is a promising target for asthma treatment, however, studies are needed to define the mechanisms by which RAGE promotes airway inflammation and asthma

Future Directions

Determine the effects of RAGE haplodeficiency on airway hyperresponsiveness by FlexiVent analysis

Determine the effects of RAGE haplodeficiency on airway mucus metaplasia and tissue inflammation

Examine which RAGE-expressing cells promote immune responses to allergens (e.g. structural or inflammatory)

Acknowledgements

This study was supported by Arrowhead Pharmaceuticals

Results

RESULTS4

DAMP and PRR

Most abundantly expressed in the lungs

Binds several ligands

Activates an array of signaling

pathways

SNPs are associated with variation in lung function

RAGE+/+

Allergen Exposure

RAGE-/-

HMGB1

IL-33

HMGB1

VCAM1 VCAM1

RAGE-deficient (heterozygous global knockout) mice are protected in several models of T2hi asthma/allergic airway

Endothelium Activation

rIL-33

TH2

ILC2

ILC2 ILC2

Endothelium Activation

rIL-33

TH2

disease (AAD)

RAGE is required at several steps in the pathologic process of T2hi AAD

Here we examine the effects of RAGE haplodeficiency in multiple models of AAD

rIL-4/5/13

IL-4

IL-13

TH2-skewing

AHR

B-cell class switching

Chemokine

Chemokine production

production

Mucus Metaplasia

IL-5

Eosinophil Proliferation Activation Recruitment

IL-5 IL-13 IL-4

rIL-4/5/13

Perkins et al. Allergy 2021

Methods

Experimental models of asthma

We utilized three established mouse models of T2hi asthma/ allergic airway disease:

Alternaria alternata

House dust mite (HDM) and

rIL-33

Allergens (Alternaria/HDM) were purchased from Greer Labs and rIL-33 from Biolegend

Animals

Wild-type (C57BL/6NTac), RAGE-Het (+/-) and RAGE-KO (-/-) were used in each experimental model

Inflammation

Bronchoalveolar lavage (BAL) cellular inflammation was determined by total cell counts and differential counts by cytospin analysis

BAL cytokines were measured by multiplex assay (Eve Technologies)

Chronic disease of the airways:

Persistent inflammation

Mucus hypersecretion

Airway hyper-reactivity

Caused by environmental and genetic factors.

Pathologically heterogeneous

Affects ~400 million people worldwide

Nearly 10% of cases are severe

Accounts for 50% of healthcare burden of asthma in the US

Half of severe asthmatics present with "Type 2-high" (T2hi) phenotype

T2hi asthma is driven by type 2 cytokines: Interleukin(IL)-4, IL-5, and IL-13

The mechanisms of T2hi asthma remain incompletely understood

Lambrecht et al. Immunity 2019

A Day 0 3 6 B C

Wild-type (+/+)

9 10

END

i.n. SAL or Alt (25µg)

RAGE-Het (+/-)

Day 0

Challenge

RAGE-KO (-/-)

i.n. SAL or HDM (50µg)

14 17 18

END

i.n. SAL or HDM (25µg)

Sensitization

Challenge

Figure 1 (A) Experimental design: Wild-type, RAGE-Het and RAGE-KO mice were challenged with models of Alternaria and HDM induced allergic airway inflammation as indicated. (B) BAL Total cells for mice challenged with Alternaria.

indicated comparison, N=3-6/group.

Wild-type (+/+)

Day 0

2 3

RAGE-Het (+/-)

END

i.n. SAL or rIL-33 (1µg)

RAGE-KO (-/-)

Challenge

Figure 2 (A) Experimental design: Wild-type, RAGE-Het and RAGE-KO mice were challenged with a model of rIL-33 induced allergic airway inflammation as indicated. (B) BAL Total cells.

BAL sRAGE levels. *P<0.05 for indicated comparison, N=3-6/group.

Figure 3 BAL type 2 cytokine levels. (A) Interleukin-4 (IL-4),

(B) Interleukin-5 (IL-5) and (C) Interleukin-13 (IL-13) *P<0.05 for indicated comparison, N=3-

6/group.

Figure 4 BAL Monocyte,

macrophage

chemokines.

(A) Monokine-induced by gamma interferon (MIG), (B) Macrophage inflammatory protein-1 (MIP-1) and (C) Macrophage inflammatory protein-1 (MIP-1). *P<0.05 for indicated comparison, N=3-6/group.

A B C

Figure 5.

cytokines.

BAL Th1/Th17

(A)

Tumor

necrosis factor  (TNF ),

(B) Granulocyte-colony stimulating factor (G-CSF) and (C) Interferon gamma induced protein 10 (IP-10)

*P<0.05 for indicated

comparison, N=3-6/group.

Asthma

Inflammation

RAGE expression

RAGE inhibition

Progressive reduction in RAGE expression correspondingly reduces airway inflammation in well established models of asthma

Disclaimer

Arrowhead Pharmaceuticals Inc. published this content on May 18, 2025, and is solely responsible for the information contained herein. Distributed via Public Technologies (PUBT), unedited and unaltered, on May 19, 2025 at 00:05 UTC.