Asprosin-neutralizing antibodies as a treatment for metabolic syndrome
- Harrington Discovery Institute, University Hospitals, United States
- Texas Therapeutics Institute, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, United States
- Baylor College of Medicine, United States
- Department of Pathology, Case Western Reserve University, United States
- Department of Medicine, Case Western Reserve University, United States
- Department of Genetics and Genome Sciences, Case Western Reserve University, United States
Figures
Figure 1 with 5 supplements
Acute asprosin neutralization reduces plasma glucose, appetite, and body weight in diet-induced obese mice.
(a–d) Cumulative food intake and body weight change (measured 24 hr post-treatment), baseline blood glucose (measured at hour 2, 4, and 6 post-mAb treatment) and plasma insulin (measured 6 hr …
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Figure 1—source data 1
Raw data and statistical analysis values for Figure 1 and Figure 1—figure supplement 1.
- https://cdn.elifesciences.org/articles/63784/elife-63784-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Higher blood glucose and insulin levels in metabolic syndrome patients are associated with elevated asprosin levels.
Blood glucose, plasma insulin, and asprosin levels were measured in metabolic syndrome male patients (n = 10; BMI > 25) and age-matched male subjects with normal BMI (<25). Asterisk (*) indicate the …
Figure 1—figure supplement 2
Asprosin neutralization does not cause glucosuria.
Urine glucose levels were measured at 0, 2, 4, and 6 hr after a single dose of anti-asprosin mAb or control, isotype-matched IgG (250 µg/mouse) in fasting 16-week-old male C57BL/6J diet-induced …
Figure 1—figure supplement 3
Acute asprosin neutralization reduces blood glucose, but not appetite and body weight in lean mice.
Cumulative food intake and body weight change (at 24 hr post-treatment), baseline blood glucose (at hour 2, 4, and 6 post-mAb treatment) and plasma insulin (at 6 hr post-treatment) was measured …
Figure 1—figure supplement 4
Blood glucose, appetite, and body weight remain unaltered in response to anti-asprosin treatment in Fbn1NPS/+ mice.
Cumulative food intake and body weight change (at 24 hr post-treatment) and baseline blood glucose (at hour 2, 4, and 6 post-mAb treatment) were measured after a single dose of anti-asprosin mAb …
Figure 1—figure supplement 5
Anti-asprosin monoclonal antibody treatment does not lead to confounding side effects.
Markers of general sickness (half hourly pedestrian activity and wheel running activity), plasma levels of pro-inflammatory (IL1β and TNFα), and anti-inflammatory (IL10 and TGFβ) cytokines, markers …
Figure 2
Asprosin neutralization corrects hyperglycemia, hyperphagia, and hypertriglyceridemia in a dose-dependent manner.
Baseline blood glucose (measured 4 hr post-treatment), plasma insulin and triglyceride (TG; at 24 hr post-treatment) levels, cumulative food intake, body weight change (measured 24 hr …
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Figure 2—source data 1
Raw data and statistical analysis values for Figure 2 and Figure 2-supplements.
- https://cdn.elifesciences.org/articles/63784/elife-63784-fig2-data1-v2.xlsx
Figure 3 with 2 supplements
Immunologic neutralization fully rescues the metabolic effects of viral induction of plasma asprosin.
(a) Body weight change was measured over 15 days after 12-week-old male C57Bl/6 mice were tail-vein-injected with Ad-empty or Ad-FBN1 (3.6 × 109 pfu/mouse, n = 12/group) viruses. Downward arrow …
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Figure 3—source data 1
Raw data and statistical analysis values for Figure 3 and figure 3-supplements.
- https://cdn.elifesciences.org/articles/63784/elife-63784-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Viral overexpression of human asprosin results in a MS-like phenotype in lean mice.
(a–d) Body weight change, cumulative food intake, blood glucose, and plasma insulin were measured 15 days after 12-week-old male C57Bl/6 mice were tail-vein-injected with Ad-empty or Ad-FBN1 (3.6 × …
Figure 3—figure supplement 2
Raw body weight values of Ad5 and AAV experiments.
Body weight was measured after 12-week-old male C57Bl/6 mice were tail-vein-injected with Ad-empty or Ad-FBN1 (3.6 × 109 pfu/mouse, n = 12/group) virus, Ad-empty or Ad-asprosin (5 × 1010 pfu/mouse, …
Figure 4 with 3 supplements
Chronic asprosin neutralization improves metabolic syndrome in three independent mouse models.
(a–d) Body weight and glucose tolerance were measured on day 11 (a, b) and day 13 (c, d) after 10 days of once daily intra-peritoneal injection of control, isotype-matched IgG, or anti-asprosin mAb …
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Figure 4—source data 1
Raw data and statistical analysis values for figure and figure-supplements.
- https://cdn.elifesciences.org/articles/63784/elife-63784-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
Raw body weight values of 14 day mAb experiments.
Body weight was measured daily with 14 days of once daily intra-peritoneal injection of control, isotype-matched IgG, or anti-asprosin mAb in 30-week-old male DIO mice (n = 8 or 9/group), …
Figure 4—figure supplement 2
Chronic asprosin neutralization reduces body weight in LepRdb/db and DIO mice.
Body weight and % change in body weight were measured after once daily intra-peritoneal injection of control, isotype-matched IgG, or anti-asprosin mAb (250 µg/day) in 30-week-old male Leprdb/db …
Figure 4—figure supplement 3
Asprosin neutralization improves dyslipidemia in MS mouse models.
(a–f) Plasma levels of total cholesterol (TC), low-density lipoproteins (LDL), high-density lipoprotein (HDL), triglycerides (TG), free fatty acids (FFA), and glycerol were measured 6 hr after …
Figure 5
A 14 day course of anti-asprosin mAb treatment reduces adipose mass in diet-induced obese mice.
Lean mass (a), body weight (b), intra peritoneal fat volume (c), and subcutaneous fat volume (d) were measured pretreatment (day 0) and post-treatment (day 14) of once daily intra-peritoneal …
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Figure 5—source data 1
Raw data and statistical analysis values for Figure 5 and figure 5-supplements.
- https://cdn.elifesciences.org/articles/63784/elife-63784-fig5-data1-v2.xlsx
Figure 6 with 1 supplement
Pharmacokinetics of epitope-agnostic anti-asprosin mAbs from different sources.
(a–c) Neutralization of asprosin using mAbs from different sources is equally protective. mAb generated using mouse immunization with a 28-mer asprosin peptide (mouse mAb), immunization of rabbits …
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Figure 6—source data 1
Raw data and statistical analysis values for figure 6 and figure-supplements.
- https://cdn.elifesciences.org/articles/63784/elife-63784-fig6-data1-v2.xlsx
Figure 6—figure supplement 1
Characterization of the asprosin-neutralizing antibodies.
Free asprosin levels were detected by ELISA against recombinant asprosin (1 nM) preincubated with various concentrations of rabbit and fully human anti-asprosin mAbs or the IgG control antibody.
Figure 7
Anti-asprosin mAbs from different sources neutralize asprosin’s orexigenic and glucogenic function by blunting AgRP+ neuronal firing and hepatic cAMP signaling.
(a–f) The anti-asprosin neutralizing antibodies reversibly inhibit asprosin’s effect on AgRP+ neuronal activity. Firing frequency (a, b) and membrane potential (d, e) of AgRP+ neurons were recorded …
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Figure 7—source data 1
Raw data and statistical analysis values for Figure 7.
- https://cdn.elifesciences.org/articles/63784/elife-63784-fig7-data1-v2.xlsx
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Animal resource (Mus musculus) | C57BL/6J mice | Jackson Laboratory | RRID:IMSR_JAX:000664 | |
| Animal resource (Mus musculus) | C57BL/6-Fbn1em1Chop/J | Jackson Laboratory | RRID:IMSR_JAX:033548 | |
| Animal resource (Mus musculus) | C57BL/6J DIO | Jackson Laboratory | RRID:IMSR_JAX:380050 | |
| Animal resource (Mus musculus) | B6.BKS(D)-Leprdb/J | Jackson Laboratory | RRID:IMSR_JAX:000697 | |
| Research rodent food | Dustless pellet diet | Bio-Ser | F0173 | |
| Research rodent food | High-fat diet | Envigo Teklad | TD.06414 | |
| Research rodent food | AMLN diet | Research Diets | D09100301 | |
| Antibody | Mouse anti-asprosin mAb | This paper | ||
| Antibody | Rabbit anti-asprosin mAb | This paper | ||
| Antibody | Human anti-asprosin mAb | This paper | ||
| Antibody | Anti-mouse secondary antibody | Tonbo Biosciences | 72–8042 M001 | 1:10,000 |
| Antibody | Anti-rabbit secondary antibody | Cytiva’s Amersham ECL | NA934-1ML | 1:10,000 |
| Adenoviral vector | Ad5-hFBN1 | This paper | 3.6 × 109 pfu/mouse | |
| Adenoviral vector | Ad5-hAsprosin | This paper | 5 × 1010 pfu/mouse | |
| Adeno-associated viral vector | AAV8-hAsprosin | This paper | 1 × 1012 GC/mouse | |
| Adenoviral vector | Ad5-Empty | This paper | 3.6 × 109 pfu/mouse or 5 × 1010 pfu/mouse | |
| Adeno-associated viral vector | AAV8-Empty | This paper | 1 × 1012 GC/mouse | |
| Commercial kit | cAMP ELISA kit | Crystal Chemicals | Catalog # 581001 | |
| Commercial kit | Glucose quantitation kit | Cayman Chemicals | Catalog # 10009582 | |
| Commercial kit | Mouse insulin ELISA kit | Crystal Chemicals | Catalog # 90080 | |
| Commercial kit | Human insulin ELISA kit | Raybiotech | Catalog # ELH-Insulin | |
| Commercial kit | IL1β ELISA kit | Abcam | Catalog # ab197742 | |
| Commercial kit | TNF ELISA kit | Abcam | Catalog # ab208348 | |
| Commercial kit | IL10 ELISA kit | Thermo Scientific | Catalog # BMS614INST | |
| Commercial kit | TGFβ ELISA kit | R and D systems | Catalog # DB100B | |
| Commercial kit | ALT ELISA kit | Abcam | Catalog # ab105134 | |
| Commercial kit | AST ELISA kit | Abcam | Catalog # ab263882 |
