Research ArticleObesity

Long-acting MIC-1/GDF15 molecules to treat obesity: Evidence from mice to monkeys

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Science Translational Medicine  18 Oct 2017:
Vol. 9, Issue 412, eaan8732
DOI: 10.1126/scitranslmed.aan8732
  • Fig. 1. GDF15 is up-regulated with obesity, and AAV-GDF15 improves metabolic parameters in DIO mice.

    (A) Microarray signal intensity of GDF15 in the liver and fat tissues from C57BL/6 and ob/ob mice (n = 5 to 6). (B) Serum GDF15 concentrations in lean and obese mice (n = 5 to 8), rats (n = 5 to 6), and humans (n = 16). (C) Body weight, average daily food intake 1 month after AAV-hGDF15 injection; glucose, insulin, triglyceride, and cholesterol concentrations 5 months after AAV injection; and serum hGDF15 concentrations 12, 81, and 350 days after AAV injection in male B6D2F1 DIO mice injected with empty vector or AAV-hGDF15 (n = 5 to 15). (D) Fat mass, lean mass, and bone mineral density of 18-month-old male B6D2F1 DIO mice injected with empty vector 12 months after AAV injection, 18-month-old male B6D2F1 DIO mice injected with AAV-hGDF15 12 months after AAV injection, and a group of untreated 12-week-old mice on normal chow (n = 3). *P < 0.05, **P < 0.01, and ***P < 0.001 versus empty vector by analysis of variance (ANOVA).

  • Fig. 2. GDF15 crystal structure and enabled design of GDF15 molecules.

    (A) Side view and (B) top view of the crystal structure of GDF15 dimer at 2.3 Å with individual subunits shown in gold and cyan. The N terminus and C terminus are highlighted by dotted spheres. (C) Close-up view of the C terminus of GDF15. One GDF15 monomer is shown in beige surface representation. The C-terminal residue Ile112 of the other monomer is shown in stick representation. The van der Waals radius of Ile112 is shown as dotted spheres. (D) Summary of the ED50 of GDF15 fusion molecules. DhFc, hinge-deleted Fc fragment; CpmFc, charge-paired mutant of Fc fragment; DhCpmFc, hinge-deleted charge-paired Fc fragment; ScFc, single-chain Fc fragment.

  • Fig. 3. Fc fusion GDF15 molecules improve metabolic parameters in obese mice and obese cynomolgus monkeys.

    (A) Body weight, OGTT glucose AUC, insulin, triglyceride, cholesterol concentrations, and food intake of male C57BL/6 DIO mice treated weekly with vehicle, rosiglitazone, or Fc fusion GDF15 proteins (0.1, 1, or 10 nmol/kg) for 5 weeks (n = 12). *P < 0.05, **P < 0.01, and ***P < 0.001 versus vehicle by ANOVA. (B and C) Cynomolgus monkeys received weekly subcutaneous doses (days 0, 7, 14, 21, 28, and 35) of vehicle (closed square; n = 10), ScFc (open triangle; n = 5), or DhCpmFc (open circle; n = 8) for 6 weeks. (B) Plasma chemistries and body weights were measured 6 days after each weekly injection before the morning meal (days 6, 13, 20, 27, 34, and 41); food intake measurements occurred daily. Data are expressed as group means ± SEM. (C) OGTT data are represented as AUC (glucose AUC, mg/dl per hour; insulin AUC, ng/ml per hour; 0 to 180 min). Cynomolgus monkeys suspected to be positive for anti-GDF15 antibodies were excluded from data analysis (five monkeys for ScFc and two for DhCpmFc). Statistical analysis was performed by analysis of covariance (ANCOVA), and statistical significance is denoted as #P < 0.05, ##P < 0.01, and ###P < 0.01 versus vehicle for ScFc; *P < 0.05, **P < 0.01, and ***P < 0.01 versus vehicle for DhCpmFc.

  • Fig. 4. GDF15 delays gastric emptying, changes food preference, and activates AP neurons.

    (A) Percentage of liquid emptied out of the stomach 5 and 15 min after oral gavage (n = 8). Vehicle or GDF15 protein was dosed 30 min before oral administration of phenol red solution. *P < 0.05, **P < 0.01, and ***P < 0.001 versus vehicle by ANOVA. (B) Percentage of normal chow or condensed milk diet consumed before or after GDF15 treatment (n = 8). *P < 0.05, **P < 0.01, and ***P < 0.001, normal chow versus condensed milk diet, by ANOVA. (C) Representative thick sections of the AP showing c-FOS–positive neurons in mice dosed with amylin or high-dose GDF15 protein 30 min after intraperitoneal injection. Scale bars, 50 μm. (D) Fc immunoreactivity in colon sections after administration of DhCpmFc or controls. Scale bars, 25 μm. Corresponding plasma concentrations are indicated below the respective images. (E) Colocalization of Fc immunoreactivity with the neuronal marker pgp9.5 in colon sections. Scale bars, 25 μm.

Supplementary Materials

  • www.sciencetranslationalmedicine.org/cgi/content/full/9/412/eaan8732/DC1

    Fig. S1. Improved metabolic parameters in AAV-hGDF15–injected ob/ob, db/db, and KKAy mice.

    Fig. S2. Effects of rmGDF15, rhGDF15, and mGDF15 antibodies on food intake in ob/ob mice.

    Fig. S3. Metabolic parameters in obese mice and obese cynomolgus monkeys treated with rhGDF15 protein.

    Fig. S4. PK of GDF15 proteins.

    Fig. S5. Model of DhCpmFc fusion protein.

    Fig. S6. PK of long-acting GDF15 proteins.

    Fig. S7. Reduced food intake and body weight in Zucker fatty rats treated with DhCpmFc protein.

    Fig. S8. No delay in gastric emptying in GDF15-treated mice after vagotomy.

    Fig. S9. Thick sections of the AP showing c-FOS–positive neurons in additional amylin and high-dose GDF15 peptide–dosed mice.

    Fig. S10. Biodistribution of Fc immunoreactivity in jejunum, stomach, and liver.

    Table S1. Pathology report of DIO mice 1 year after control AAV or AAV-GDF15 injection.

    Table S2. Statistics of crystallographic data and refinement.

    Table S3. Individual subject-level data of quantitative studies (provided as an Excel file).

  • Supplementary Material for:

    Long-acting MIC-1/GDF15 molecules to treat obesity: Evidence from mice to monkeys

    Yumei Xiong, Kenneth Walker, Xiaoshan Min, Clarence Hale, Thanhvien Tran, Renee Komorowski, Jerry Yang, Jasmine Davda, Noi Nuanmanee, Dao Kemp, Xiaozhen Wang, Hantao Liu, Silke Miller, Ki Jeong Lee, Zhulun Wang, Murielle M. Véniant*

    *Corresponding author. Email: mveniant{at}amgen.com

    Published 18 October 2017, Sci. Transl. Med. 9, eaan8732 (2017)
    DOI: 10.1126/scitranslmed.aan8732

    This PDF file includes:

    • Fig. S1. Improved metabolic parameters in AAV-hGDF15–injected ob/ob, db/db, and KKAy mice.
    • Fig. S2. Effects of rmGDF15, rhGDF15, and mGDF15 antibodies on food intake in ob/ob mice.
    • Fig. S3. Metabolic parameters in obese mice and obese cynomolgus monkeys treated with rhGDF15 protein.
    • Fig. S4. PK of GDF15 proteins.
    • Fig. S5. Model of DhCpmFc fusion protein.
    • Fig. S6. PK of long-acting GDF15 proteins.
    • Fig. S7. Reduced food intake and body weight in Zucker fatty rats treated with DhCpmFc protein.
    • Fig. S8. No delay in gastric emptying in GDF15-treated mice after vagotomy.
    • Fig. S9. Thick sections of the AP showing c-FOS–positive neurons in additional amylin and high-dose GDF15 peptide–dosed mice.
    • Fig. S10. Biodistribution of Fc immunoreactivity in jejunum, stomach, and liver.
    • Table S1. Pathology report of DIO mice 1 year after control AAV or AAVGDF15 injection.
    • Table S2. Statistics of crystallographic data and refinement.

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    Other Supplementary Material for this manuscript includes the following:

    • Table S3. Individual subject-level data of quantitative studies (provided as an Excel file).

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