Supplementary Information: Figures 1-6 and Table 1 RNAi-Mediated Gene Silencing in Non-Human Primate Zimmermann, T.S. et al.

Similar documents
Phase 1 Trial of ALN-VSP in Cancers Involving the Liver. Annual Meeting of the Controlled Release Society August 2, 2011

Pair-fed % inkt cells 0.5. EtOH 0.0

Spleen. mlns. E Spleen 4.1. mlns. Spleen. mlns. Mock 17. Mock CD8 HIV-1 CD38 HLA-DR. Ki67. Spleen. Spleen. mlns. Cheng et al. Fig.

Development of an RNA Interference Therapeutic Targeting Angiopoietin-Like Protein 3 for Treatment of Hyperlipidemia

Supplementary Figure 1: Additional metabolic parameters of obesity mouse models and controls. (a) Body weight, (b) blood glucose and (c) insulin

AsiaTIDES 2012: Formulation and Delivery of Peptides and Oligonucleotides Strategies for Delivery of RNAi Therapeutics. March 1, 2012 Akin Akinc, PhD

2011 ASH Annual Meeting Targeting the Hepcidin Pathway with RNAi Therapeutics for the Treatment of Anemia. December 12, 2011

Supplementary Information

Supplementary figure 1. Systemic delivery of anti-cd47 antibody controls tumor growth in

Translatability of cytokine data: from animals to humans. Marie-Soleil Piche, PhD Associate Scientific Director of Immunology Charles River, Montreal

Ct=28.4 WAT 92.6% Hepatic CE (mg/g) P=3.6x10-08 Plasma Cholesterol (mg/dl)

Supplementary Figure 1. Characterization of basophils after reconstitution of SCID mice

Supplementary Figure 1. DNA methylation of the adiponectin promoter R1, Pparg2, and Tnfa promoter in adipocytes is not affected by obesity.

Electron micrograph of phosphotungstanic acid-stained exosomes derived from murine

Supplementary Figure 1 IMQ-Induced Mouse Model of Psoriasis. IMQ cream was

Supplementary Figure 1. Western blot of hippocampal lysates from WT and Adcy1 KO mice demonstrates the specificity of the ADCY1 antibody.

Table S1. Quantitative RT-PCR primers

PCSK9 RNAi Therapeutics. Kevin FitzGerald

ECM1 controls T H 2 cell egress from lymph nodes through re-expression of S1P 1

Supplement Figure S1. Real Time PCR analysis of mrna levels of C/EBPα and PU.1 in wild type (WT) and NQO1-null (NQO1-/-) mice.

complemented with SipA ( SipA/pSipA) or SL1344 WT for 48 hours, after which the

Supplemental Materials for. Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to. FTY720 during neuroinflammation

Supplementary Figure 1. Deletion of Smad3 prevents B16F10 melanoma invasion and metastasis in a mouse s.c. tumor model.

Nature Immunology: doi: /ni Supplementary Figure 1. Cytokine pattern in skin in response to urushiol.

Therapeutic Efficacy of a TLR7 Agonist for HBV Chronic Infection in Chimpanzees

Supplementary Information

NKTR-255: Accessing The Immunotherapeutic Potential Of IL-15 for NK Cell Therapies

Supplementary Figure 1. AdipoR1 silencing and overexpression controls. (a) Representative blots (upper and lower panels) showing the AdipoR1 protein

Stacey Melquist #AHA16

MATERIALS AND METHODS. Neutralizing antibodies specific to mouse Dll1, Dll4, J1 and J2 were prepared as described. 1,2 All

O CYP1B1 CYP1A1 +DNA meh HO OH H NO 2. N OR NATs NQO1 N OH. SULTs

Detection and significance of PD-1.3 SNP (rs ) and IL28B SNP (rs ) in patients with current or past hepatitis B virus (HBV) infection

SUPPLEMENTARY INFORMATION

Supplementary Figure 1. Expression of CUGBP1 in non-parenchymal liver cells treated with TGF-β

Supplementary Figure 1. Antibiotic partially rescues mice from sepsis. (ab) BALB/c mice under CLP were treated with antibiotic or PBS.

The levels of mrna expression in the mouse brain were measured at 52 dpi after the brains were divided into five regions: cortex, hippocampus,

Supplementary Figure 1. Repression of hepcidin expression in the liver of mice treated with

Supplementary Figure S1. PTPN2 levels are not altered in proliferating CD8+ T cells. Lymph node (LN) CD8+ T cells from C57BL/6 mice were stained with

pplementary Figur Supplementary Figure 1. a.

Phenomena first observed in petunia

PK and PD Properties of Antisense Oligonucleotides: Bridging Nonclinical to Clinical

Supplementary Figure 1

Supplemental Table 1. Primers used for RT-PCR analysis of inflammatory cytokines Gene Primer Sequence

Supplementary Figures

Supplementary Figures

Supplementary table I. Real-time primers used in the study. The fold change was obtained by

John Langowski, Ph.D. Nektar Therapeutics San Francisco, CA USA

NKTR-255: Accessing IL-15 Therapeutic Potential through Robust and Sustained Engagement of Innate and Adaptive Immunity

Supplementary Figure 1.

RNA interference induced hepatotoxicity results from loss of the first synthesized isoform of microrna-122 in mice

Nature Medicine: doi: /nm.4324

Supplemental Table I.

Supplementary Figure (OH) 22 nanoparticles did not affect cell viability and apoposis. MDA-MB-231, MCF-7, MCF-10A and BT549 cells were

ACC ELOVL MCAD. CPT1α 1.5 *** 0.5. Reverbα *** *** 0.5. Fasted. Refed

ALN-PCSsc, an RNAi Investigational Agent That Inhibits PCSK9 Synthesis With the Potential for Effective Bi-Annual Dosing: Interim Results

Supplementary Information

Supplementary Figure 1. Generation of knockin mice expressing L-selectinN138G. (a) Schematics of the Sellg allele (top), the targeting vector, the

Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained

LNA-mediated silencing of microrna-122 in African green monkeys

Doctor of Philosophy

Supporting Information Table of Contents

Reduction of metastatic and angiogenic potency of malignant cancer by Eupatorium. fortunei via suppression of MMP-9 activity and VEGF production

Supplementary fig. 1. Crystals induce necroptosis does not involve caspases, TNF receptor or NLRP3. A. Mouse tubular epithelial cells were pretreated

Nature Immunology: doi: /ni Supplementary Figure 1. Gene expression profile of CD4 + T cells and CTL responses in Bcl6-deficient mice.

Nature Immunology doi: /ni Supplementary Figure 1. Raf-1 inhibition does not affect TLR4-induced type I IFN responses.

Genome-editing via Oviductal Nucleic Acids Delivery (GONAD) system: a novel microinjection-independent genome engineering method in mice

Regulation of T cell proliferation by JMJD6 and PDGF-BB during chronic. hepatitis B infection

Online Appendix Material and Methods: Pancreatic RNA isolation and quantitative real-time (q)rt-pcr. Mice were fasted overnight and killed 1 hour (h)

Supplementary Figure 1: Hsp60 / IEC mice are embryonically lethal (A) Light microscopic pictures show mouse embryos at developmental stage E12.

Protection against doxorubicin-induced myocardial dysfunction in mice by cardiac-specific expression of carboxyl terminus of hsp70-interacting protein

Cystinosis Research Foundation Progress Report. Energy homeostasis and muscle wasting in nephropathic cystinosis. Final Report

Supplementary Figure 1 Binding of PAR1-RIP to (A) anionic liposomes consisting of phosphatidylserine and (B) zwitterionic liposomes composed of

Supplementary Figure 1. mrna expression of chitinase and chitinase-like protein in splenic immune cells. Each splenic immune cell population was

Supporting Information

GFP/Iba1/GFAP. Brain. Liver. Kidney. Lung. Hoechst/Iba1/TLR9!

Supplementary Table 1. Table showing different gene specific primers used in real-time PCR.

Supplemental Material. Results

Suppl Video: Tumor cells (green) and monocytes (white) are seeded on a confluent endothelial

Type of file: PDF Size of file: 0 KB Title of file for HTML: Supplementary Information Description: Supplementary Figures

Alnylam RNAi Roundtable. May 13, 2011

Stability of VACUETTE Lithium Heparin Separator tubes with modified centrifugation conditions

Prurisol: A New Small Molecule under investigation for the treatment of Psoriasis

Supplementary Figure S1. Effect of Glucose on Energy Balance in WT and KHK A/C KO

Supplementary Materials and Methods

Supplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured

SUPPLEMENTARY INFORMATION

Social deficits in Shank3-deficient mouse models of autism are rescued by histone deacetylase (HDAC) inhibition

Supplementary methods:

IL-6Rα IL-6RαT-KO KO. IL-6Rα f/f bp. f/f 628 bp deleted 368 bp. 500 bp

Supplementary Fig. 1. Delivery of mirnas via Red Fluorescent Protein.

Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, Boston, MA 02115, USA 2

BALANCE BETWEEN ESTROGENS AND PROINFLAMMATORY CYTOKINES REGULATES CHEMOKINE PRODUCTION INVOLVED IN THYMIC GERMINAL CENTER FORMATION

Serum cytokine levels in control and tumor-bearing male and female mice at day 15.

VITROS MicroSlide Assay Summary

Supplementary Figure 1: si-craf but not si-braf sensitizes tumor cells to radiation.

AdPLA ablation increases lipolysis and prevents obesity induced by high fat feeding or leptin deficiency

Nature Immunology: doi: /ni Supplementary Figure 1. Id2 and Id3 define polyclonal T H 1 and T FH cell subsets.

Pharmaceutical Drug Development Consulting

Supplementary Figure 1

Fig. S1. REGN1500 reduces plasma levels of cholesterol, TG and NEFA in WT and Ldlr -/- mice. (A) WT

Transcription:

Supplementary Information: Figures 1-6 and Table 1 RNAi-Mediated Gene Silencing in Non-Human Primate Zimmermann, T.S. et al.

a. 12 14 Relative apob mrna (%) 8 6 4 2 Relative apob mrna (%) 12 8 6 4 2 5 25 12.5 Dose (mg/kg) 1.5.25.1 Dose (mg/kg) Supplementary Figure 1 Comparison of Chol-siApoB-1 to SNALP formulated siapob-1 activity in mice. a, b, Dose-dependent silencing of liver apob mrna after administration of either Chol-siApoB-1 (a) or SNALP siapob-1 (b). Liver apob mrna levels were quantified relative to GAPDH mrna three days after i.v. administration of sirna. Data are mean values relative to the saline treatment group ± s.d. Chol-siApoB- 1 was administered at doses of, 5, 25, or 12.5 mg/kg (n = 6 per group) (a) and SNALP siapob-1 was administered at sirna doses of 1,.5,.25 and.1 mg/kg (n = 4 per group) (b). 12 %Injected Dose 8 6 4 Sense Antisense 2.25.5 1 2 4 6 8 12 Time (h) Supplementary Figure 2. Pharmacokinetics of SNALP-formulated siapob-2 in cynomolgus monkeys. Plasma clearance over a 24 h period of the sense and antisense strands of siapob-2 in animals treated with 2.5 mg/kg SNALP siapob-2 (n = 2). The time course is shown extending to 12 h as no sirna was detected at 24 h. Each data point represents the group mean ± s.d. of the percent injected dose.

a. % Injected Dose 1 1 % Injected Dose 1 1.1.1 4 8 12 16 2 2 4 Time (h).1 Liver Spleen Lungs Kidney Heart Femur Thymus Small intestine Large intestine Muscle Fat Supplementary Figure 3. Pharmacokinetics and biodistribution of SNALP in mice. a, Plasma clearance and b, Tissue biodistribution of 3 H-CHE-labeled SNALP in BALB/c mice. Each mouse received a single i.v. injection of 2 mg/kg SNALP-formulated siapob-mm via the tail vein. Tissue biodistribution was analyzed 24 h after treatment. Data are expressed as mean ± s.d (n = 4). sirna: 5 4 3 2 1 LL1 LL2 LL3 M1 M2 M3 RL1 RL2 RL3 C1 C2 C3 LL1 LL2 LL3 M1 M2 M3 RL1 RL2 RL3 C1 C2 C3 LL1 LL2 LL3 M1 M2 M3 RL1 RL2 RL3 C1 C2 C3 ApoB/GAPDH mrna SNALP siapob-2 cyno-1 SNALP siapob-2 cyno-2 Supplementary Figure 4. Uniform biodistribution and silencing activity of SNALPsiApoB-2 in liver. ApoB mrna and siapob-2 accumulation were quantitated in 12 separate liver biopsy samples collected from the left lateral lobe (LL), median lobe (M), right lateral lobe (RL) and caudate lobe (C) 24 h after animals were treated with either saline or 2.5 mg/kg SNALP siapob-2. ApoB mrna was quantified relative to GAPDH mrna. Data are mean values for three measurements per liver biopsy ± s.d.

a. c. sirna Cleavage Site siapob-2 Cleavage Site apob mrna 5 uagaagggaaucuuauauuugauccaaataa UCCCUUAGAAUAUAAACUAGGUU-5 siapob-2 AS RNA Adaptor GR5 GR5N ApoB Rev2 ApoB Rev1 ApoB GSP 3' wt apob mrna Sequence SNALP siapob-2 Cyno - 1 Cyno - 2 2.5 mg/kg Adaptor sirna Cleavage Site 75 5 25 Supplementary Figure 5. Confirmation of RNAi-mediated gene silencing in nonhuman primates. a-c, 5 -RACE analysis demonstrating that silencing of apob mrna is due to RNAi-mediated mrna cleavage. (a) Scheme depicting the position of the predicted siapob-2 cleavage site relative to nested primers used for PCR amplification of the cleavage fragment. (b) Agarose gel of 5 RACE-PCR amplification product showing the predicted product of RNAi in the liver of SNALP siapob-2 treated animals and not in a saline-treated control animal. (c) Sequencing chromatograms of 5 -RACE PCR amplification products. The predicted site of siapob-2 mediated cleavage of apob mrna, 1 nucleotides from the 5 end of the antisense strand of siapob-2, is illustrated at top.

a. 4. 2.5 3.5 3. 2. apob/gapdh 2.5 2. 1.5 apob/gapdh 1.5 1. 1..5.5 A1 A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 2.5 mg/kg 1 mg/kg A5 A6 B5 B6 C5 C6 2.5 mg/kg 1 mg/kg Supplementary Figure 6. Silencing of jejunum apob mrna was not observed. a, b, ApoB mrna levels for eight tissue sections isolated from the jejunum were quantified relative to GAPDH mrna for each animal. Data are mean values ± s.d. Jejunum apob mrna levels 48 h after treatment (n = 4) (a) and 11 d after treatment (n = 2) (b).

Supplementary Table 1 a, ALT, AST, total bilirubin and urea nitrogen levels for saline and SNALP-treated cynomolgus monkeys.* ALT (U/l) AST (U/l) Bilirubin (mg/dl) Urea Nitrogen (mg/dl) Pre-dose 52 ± 12 69 ± 15.38 ±.8 22 ± 3 24 h 58 ± 22 72 ± 29.33 ±.8 19 ± 3 48 h 52 ± 19 57 ± 18.4 ±.9 19 ± 3 144 h 36 ± 13 47 ± 18.35 ±.7 21 ± 1 264 h 34 ± 9 39 ± 5.3 ±. 24 ± 1 1. mg/kg Pre-dose 5 ± 15 58 ± 13.35 ±.8 21 ± 6 24 h 47± 16 57 ± 27.28 ±.4 19 ± 2 48 h 48 ± 18 45 ± 2.3 ±. 19 ± 3 144 h 51 ± 21 48 ± 13.35 ±.7 22 ± 1 264 h 43 ± 7 38 ± 4.25 ±.7 21 ± 3 2.5 mg/kg Pre-dose 47 ± 14 75 ± 24.33 ±.5 22 ± 3 24 h 152 ± 196 123 ± 128.32 ±.4 2 ± 3 48 h 1167 ± 1157 1555 ± 1727.43 ±.15 21 ± 2 144 h 582 ± 32 53 ± 16.3 ±. 2 ± 6 264 h 151 ± 56 45 ± 13.35 ±.7 24 ± 1 *Values reported are the mean ± s.d. for each group. Pre-dose, 24 and 48 h time points had a group size of six and 144 and 264 h time points had a group size of two.

b, APTT, complement Bb, CH5, IL-6 and IFN-γ levels for saline and SNALP-treated cynomolgus monkeys. APTT (sec)* Complement Bb (µg/ml)* CH5 (U/ml)* IL-6 (pg/ml)** IFN-γ (IU/ml)** Pre-dose 26.8 ± 2.3.45 ±.6 31.83 ± 5.9 48.1 159.9.25 h 25.7 ± 2.4.53 ±.7 279.67 ± 52.66 6 h 4.7 99.9 24 h 35. 115.2 48 h 25.2 ± 3.6.46 ±.8 341.33 ± 89.1 144 h 25.9 ± 1.6 264 h 25.6 ± 4.1 1. mg/kg Pre-dose 24.6 ± 1.5.49 ±.17 293.5 ± 36.37.25 h 24.2 ± 1.5.77 ±.34 252.83 ± 29.42 6 h 24 h 48 h 24.3 ± 2..6 ±.25 311.67 ± 24.69 144 h 28.2 ±. 264 h 25.3 ±.5 2.5 mg/kg Pre-dose 27. ± 3.2.76 ±.55 278.17 ± 65.31 73.4 ± 24.3 89.5.25 h 27. ± 3..96 ±.56 233.5 ± 57.33 6 h 49.9 ± 11.3 12.1 24 h 33.4 ± 33.4 86.4 48 h 33.3 ± 4.6 1.82 ±.58 276.5 ± 57.19 144 h 27.9 ± 6.2 264 h 25.7 ± 4.2 *Values reported for APTT, complement Bb and CH5 are the mean ± s.d for each treatment group. Predose,.25 and 48 h time points had a group size of six and 144 and 264 h time points had a group size of two. **Values reported for the cytokines, IL-6 and IFN-γ, are derived from one saline animal or two 2.5 mg/kg treated animals (mean ± s.d.) in the case of IL6 and only one animal for each treatment for IFN-γ.

2024 © healthdocbox.com Privacy Policy | Terms of Service | Feedback