Pg. 1Confidential

Restoring Biological Harmony for Patients with Debilitating DiseaseRestoring Biological Harmony for Patients with Debilitating Disease

RNA Based Therapeutics as a Modality for the Effective Treatment of Multiple Fibrotic ConditionsCorrie Gallant-Behm, PhD, MQARSAnti-Fibrotic Drug Development Summit, November 14th, 2017, Boston, MA

Pg. 2

microRNA Therapeutics Regulate Systems Biology to Modify Disease

microRNA-targeted therapy is focused on disease modification by restoring homeostasis to dysregulated processes

microRNAs regulate complex biological systems

microRNA-targeted therapies are intrinsically focused on disease-relevant pathways

microRNA therapeutics particularly suited for complex, multigenic disorders

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miR-29 Pathways and Systems Control

Growth factors

Collagen transcription/translation

Post-translational modification& triple helix formation

N- and C-terminal cleavage& secretion

Fibril cross-linking

Mature collagen fibrils

miR-29

Inflammation

TGF-β + Diseased ECM

TGF-β2, TGF-β3, EGF, IGF2, IGFBP5, PDGFA, PDGFC

COL1A1, 1A2, 3A1, 5A1, 5A2, 5A3, 6A4, 6A5, 6A6, 8A1, 8A2, 9A1, 11A1, 12A1, 14A1, 22A1, 28A1

HSP47, P4HA2, P4HA3, PLOD2

PCOLCE2

LOXL2

in vivo Validated Targets

Pg. 4

miR-29 Pathways and Systems Control

MRG-201(promiR-29)

Growth factors

Collagen transcription/translation

Post-translational modification& triple helix formation

N- and C-terminal cleavage& secretion

Fibril cross-linking

Mature collagen fibrils

TGF-β2, TGF-β3, EGF, IGF2, IGFBP5, PDGFA, PDGFC

COL1A1, 1A2, 3A1, 5A1, 5A2, 5A3, 6A4, 6A5, 6A6, 8A1, 8A2, 9A1, 11A1, 12A1, 14A1, 22A1, 28A1

HSP47, P4HA2, P4HA3, PLOD2

PCOLCE2

LOXL2

in vivo Validated Targets

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A miR-29 Positive Feedback Loop in Fibrosis

Homeostasis

Matrix

TGFβ, PDGF EGFIGF

miR-29

Injury

Scar Formation

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Therapeutic Hypothesis

Homeostasis

Matrix

TGFβ, PDGF EGFIGF

promiR-29

MRG-201 restores homeostasis by modulating the positive feedback loops that maintain the fibrotic phenotype

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Fibrotic Diseases in Which Reduced miR-29 Has Been Implicated

Keloid scar.

Hypertrophic scar.

Scleroderma.

Cardiac fibrosis.

Pulmonary fibrosis – IPF, CTD-associated.

Liver fibrosis – cirrhosis, NASH, viral.

Kidney fibrosis – diabetic nephropathy, IgA.

Retinal fibrosis.

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Downregulation of miR-29 and Overexpression of miR-29 Target Genes in Keloids and Skin Scars

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miR-29 as a Therapeutic in Cutaneous Fibrosis

Preclinical models mPoC Human VolunteerWound Repair

Safety, PK,Target Engagement (PD)

PlaceboDrug Cutaneous Scleroderma

Keloids

Hypertrophic Scars

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Clinical Trial MRG201-30-001Part A: Establish Kinetics of PD Gene Expression

Incision

Day 1 Day 9 Day 16

Biopsy Biopsy RNA

miR-29qPCR

Nanostring

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miR-29 Expression Decreases and miR-29 Direct Target Expression Increases Following Incision

miR-29 QuantityGenes significantly regulated in

incised vs. intact skin (BH p-value <0.01)

Unincised Day 9Incision

Day 16Incision

0

2000

4000

6000

8000

10000m

iR-2

9 Co

py N

umbe

r Per

Cel

lns

0.002

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Parts B/C/D: Safety, Tolerability and PK in Intact Skin and Skin Incisions, PD target engagement Normal healthy volunteers Single and multiple ascending dose (N=3/cohort) 0.5-14 mg MRG-201 injected intradermally

Part B: Intact skin Safety/tolerability only, no biopsies

Part C: Incised skin, single administration Biopsy 24h after treatment for PD

Part D: Incised skin, multiple administration (6 doses over 2 weeks) Biopsy 24h after final administration for PD

RESULTS Dosing and follow-up complete All doses well tolerated up to the maximally deliverable dose No significant injection site reactions Low systemic exposure of full length MRG-201 for all subjects with many samples below limit of quantitation Treated skin biopsy biodistribution assessments (dual probe hybridization assay) showed concentrations ranging from 12 to 200 μg/g

tissue Pharmacodynamic biomarker data demonstrate target engagement with single and multiple administration of MRG-201

Line or Incision

Line orIncision

PlaceboDrug

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MRG-201 Mechanistic Proof-of-Concept in Human Incised SkinSingle Ascending Dose Cohorts C1-C3

Evidence of PD activity (mPoC) after single administration of MRG-201

Validation of preclinical PD biomarkers in human incised skin

SINGLE DOSE

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MRG-201 Treatment Significantly Blunts Fibroplasia in Human Incised SkinMultiple Ascending Dose Cohorts D1-D3

Dep

th/W

idth

(mm

) or A

rea

(mm

2 )

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MRG-201 Target Engagement Corresponds to Impact on FibroplasiaMultiple Ascending Dose Cohorts D1-D3

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miR-29 as a Therapeutic in Cutaneous Fibrosis

Preclinical models mPoC Human VolunteerWound Repair

Safety, PK,Target Engagement (PD)

PlaceboDrug Cutaneous Scleroderma

Keloids

Hypertrophic Scars

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MRG-201 Pharmacodyamic Biomarkers Translate to Multiple Fibrotic Indications

MRG-201 antimiR-29

Mouse skinvs. vehicle

Day 9 Day 16

Human skin incisionvs. unwounded

SSc skin SSc lung IPF lung

Human disease tissuevs. normal tissue

TGFB2Nedd4lPrickle1Faim2COL5A3Gimap7Cacna1gColec11ELNMfap2COL5A2COL1A1COL3A1COL11A1TGFB3Fstl1COL1A2Cytl1MMP2Sdc4Snx27Itga3NumbLbr

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MRG-201 Mimic Represses Collagen Expression in Human IPF Fibroblasts and Epithelial cells

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Day 4

125mpki.v.

Systemically Delivered MRG-201 Accumulates in the Lung

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Systemic Delivery of MRG-201 Blocks Bleomycin-induced Pulmonary Fibrosis

0 day

Bleo Takedown

10

miR-29 mimic (MRG-201)/control

3 14

Pg. 21

Aerosolized MRG-201 via Scireq Attenuates Fibrosis Induced by Bleomycin

Control

Saline

Hydr

oxyp

rolin

eµg

/rig

ht lu

ng

MRG-201 Saline

Bleomycin

0

50

100

150

200

250

ControlMRG-201 Saline

Performed at Yale w/ Naftali Kaminski

*

**

*p<0.05

0 day

Bleo Takedown

10

miR-29 mimic (MRG-201)/control

18

Pg. 22

Inhaled Pulmonary Delivery Assessed with Lovelace Respiratory Research Institute

Log2

(Fol

d Ch

ange

)

Col1a1Col1a2Col3a1ElnTgfb3Mfap2Igf1

Col

1a1

Col

1a2

Col

3a1

Eln

Tgfb

3M

fap2 Igf1

Saline/Saline Bleo/Saline0

2

4

6

Lung day 7

Col1a1Col1a2Col3a1ElnTgfb3Mfap2Igf1

0 day

Bleo Takedown

7

miR-29 mimic (MRG-201)/control

3

Pg. 23

Conclusions

MRG-201 represses expression of genes associated with extracellular matrix production

MRG-201 PD biomarker genes are conserved across species, tissues

Endogenous miR-29 and MRG-201 PD biomarker genes are inversely expressed in cutaneous incisions in normal healthy volunteers, indicating a role for miR-29 in fibrosis development/ progression

MRG-201 regulates PD biomarkers and reduces fibroplasia in skin incisions, demonstrating mechanistic proof-of-concept, and highlighting its therapeutic potential in reducing cutaneous fibrosis such as keloids, hypertrophic scars, cutaneous scleroderma

MRG-201 regulates PD biomarkers in multiple relevant lung cell types and in vivo in multiple IPF models following systemic and inhaled delivery, indicating therapeutic potential in additional fibrotic conditions such as IPF, SSC-ILD, RA-ILD

Pg. 24

Acknowledgements

miRagen: Aimee Jackson Rusty Montgomery Linda Pestano Christina Dalby Joe Piper Josh Lynch Paul Latimer Christianna Stack Kathryn Robinson

Yale: Guoying Yu Naftali Kaminski

Grants: NK and GY:

NIH Y01HL108642 NIH R01HL095397

NIH CADET II 1UH2HL123886-01

Contact: cbehm@miragen.com