Remodeling of dermal adipose tissue alleviates cutaneous toxicity induced by anti-EGFR therapy
- School of Biomedical Engineering, Shanghai Jiao Tong University, China
- Department of Dermatology, Shanghai East Hospital, Tongji University, China
Figures
Figure 1 with 4 supplements
The response of dWAT to EGFR inhibition.
(A) Representative images of the rat rash model. (B–D) H&E (B), Oil Red (C), and Caveolin-1 (D) staining of skin from control and Afa-treated rats. Scale bars: 200 μm, 500 μm and 130 μm (top to …
Figure 1—figure supplement 1
Characteristics of rat rash model.
(A) Body weight change of control and Afa-treated rats. n = 8 per group. (B) Food intake per rat of different Afa dosage. (C) Percentages of different rash grade during Afa treatment. n = 39. (D) …
Figure 1—figure supplement 2
Shaving effects rash occurrence and progress.
The back hair on the left back side was shaved while the right side was unshaved at the beginning of Afa treatment, rats were sacrificed according to the rash process of the shaved area. After …
Figure 1—figure supplement 3
Characteristics of sWAT during EGFR inhibition.
(A) Percentage of BW change of sWAT, iWAT and BAT. sWAT was obtained under the same shaved-skin area. n = 3–5 per group. (B) H&E staining of subcutaneous white adipose tissue from Ctrl and …
Figure 1—figure supplement 4
Reduced expression of PPARγ and Perilinpin-2 after Afa treatment.
(A) PPARγ and Perilinpin-2 staining of skin from Ctrl and Afa-treated rats at day 10. Scale bar: 100 μm. (B) Quantification of positive signal intensity. Data are presented as the means ± SEM. **p < …
Figure 2 with 1 supplement
Dedifferentiation of mature dermal adipocytes upon EGFR inhibition.
(A) Masson’s trichrome staining of skin sections from rats at the indicated times. n = 3–5 per group. epi refers to epidermis. Scale bars: 500 μm. (B) Changes in the thickness of the dermis in Ctrl- …
Figure 2—figure supplement 1
Additional dedifferentiation changes of dWAT and sWAT.
(A and B) mRNA levels of pro-fibrotic and pro-adipogenic genes in sWAT at one day before, and one day after rash, n = 3 per group. Data was normalized to Ctrl. Dotted line represented the mean gene …
Figure 3 with 1 supplement
EGFR inhibition blocks adipocyte progenitor differentiation.
(A) dFB cells were isolated from P1 rat skin and then cultured with adipocyte differentiation medium for 9 days in the presence or absence of Afa (10 nM) or Rosi (5 μM). Lipid production was …
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Figure 3—source data 1
Original western blot data for Figure 3D.
- https://cdn.elifesciences.org/articles/72443/elife-72443-fig3-data1-v1.zip
Figure 3—figure supplement 1
Expression levels of inflammatory receptors during dFB differentiation.
(A) Viability of dFB cells under different Afa concentrations. (B) mRNA levels of IL4R, CCR2, IL6R, TNFR, and TLR2-3 of differentiating dFB with Afa (10 nM) or Rosi (5 μM) treatment. n = 4 per …
Figure 4 with 1 supplement
Rosiglitazone alleviates lipolysis and inflammatory response.
(A) Relative expression of the Atgl and Lipe (Hsl) mRNAs in isolated dWAT on day 5. n = 3 per group. (B) Western blot images and quantifications of lipase level from isolated dWAT cells at Grade 1. …
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Figure 4—source data 1
Original western blot data for Figure 4B.
- https://cdn.elifesciences.org/articles/72443/elife-72443-fig4-data1-v1.zip
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Figure 4—source data 2
Original western blot data for Figure 4F.
- https://cdn.elifesciences.org/articles/72443/elife-72443-fig4-data2-v1.zip
Figure 4—figure supplement 1
Lipolytic effects of direct EGFRI stimulation on adipocytes.
(A) Glycerol concentration after 2 hr ISO or Afa treatment on isolated sWAT at indicated times. n = 3–5. (B and C) Lipid mass spectrometry quantification of medium- and long-free fatty acids. n = 6 …
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Figure 4—figure supplement 1—source data 1
Original western blot data for Figure 4—figure supplement 1G.
- https://cdn.elifesciences.org/articles/72443/elife-72443-fig4-figsupp1-data1-v1.zip
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Figure 4—figure supplement 1—source data 2
Original western blot data for Figure 4—figure supplement 1M.
- https://cdn.elifesciences.org/articles/72443/elife-72443-fig4-figsupp1-data2-v1.zip
Figure 5 with 2 supplements
HFD-induced dWAT expansion ameliorates rash phenotypes.
(A) Schematic of the strategy used to expand dWAT by the short-term administration of a HFD. (B) H&E staining of skin from normal diet- and HFD-fed rats. Scale bars: 600 μm. (C) Size and thickness …
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Figure 5—source data 1
Original western blot data for Figure 5F.
- https://cdn.elifesciences.org/articles/72443/elife-72443-fig5-data1-v1.zip
Figure 5—figure supplement 1
Additional characterization of HFD rats.
(A) Serum lipids of Ctrl and HFD rats. APO-B refers to apolipoprotein B, HDL-C refers to high-density lipoproteincholesterol, LDL-C refers to low-density lipoproteincholesterol, TC refers to total …
Figure 5—figure supplement 2
DCA-induced dWAT ablation aggravates rash phenotypes.
(A) Schematic of the strategy to ablate dWAT by repeatedly DCA intradermal injection. (B) H&E staining of skin from Vehicle and DCA rats. Scale bars: 300 μm. (C and D) Size (C) and thickness (D) of …
Figure 6 with 3 supplements
Prophylactic application of Rosi prevents skin toxicities.
(A) Representative photos of rash from Vehicle- and Rosi-treated rats. (B) Rash grade. (C) Rash occurrence. (D) Body weight change. (E) H&E staining of skin biopsies. Scale bars: 200 μm. (F) Number …
Figure 6—figure supplement 1
Additional effects of Rosi prevention in Afa-treated rats.
(A) PPARγ, Perilipin-2 staining of skin biopsies from Ctrl, Afa-Vehicle, Afa-Rosi rats at day 10. Scale bar: 100 μm. (B) Western-blotting images and quantifications of lipases in dFB-derived …
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Figure 6—figure supplement 1—source data 1
Original western blot data for Figure 6—figure supplement 1B.
- https://cdn.elifesciences.org/articles/72443/elife-72443-fig6-figsupp1-data1-v1.zip
Figure 6—figure supplement 2
Therapeutic application of Rosi ameliorates skin rash.
After oral gavage of Afa, Vehicle or Rosi gel was applied topically after Grade one rash occurred. (A) Representative photos of rash from Vehicle- and Rosi-treated rats. (B) H&E, Oil Red and …
Figure 6—figure supplement 3
Rosi did not interfere the anti-tumor effect of EGFRI.
Ctrl: mice gavaged with solvent. Afa 30 mpk (mg/kg): Mice were gavaged with Afa and topically administered vehicle gel. Afa 30 mpk +Rosi: Mice gavaged with Afa and topically administered Rosi gel. (A…
Figure 7
dWAT contributes to skin homeostasis during anti-EGFR therapy.
Reduction of dWAT is a hallmark of EGFRi-induced cutaneous toxicity. EGFR inhibition blocks adipocyte progenitor differentiation and induces dedifferentiation of mature dermal adipocytes. In …
Author response image 1
C18 concentrations in skin tissues from Ctrl and Afa groups after 3-day treatment.
n=3.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (Homo sapiens) | HaCaT | Cell Research | Cat. #: ZQ0044 | |
| Cell line (Homo sapiens) | PC-9 | Cell Bank of Chinese Academy of Sciences | Cat. #: SCSP-5085CSTR:19375.09.3101HUMSCSP5085 | |
| Cell line (Homo sapiens) | THP-1 | Cell Bank of Chinese Academy of Sciences | Cat. #: TCHu 57CSTR:19375.09.3101HUMTCHu57 | |
| Antibody | Rabbit monoclonal anti-ATGL | Abcam | Cat. #: ab109251 | WB (1:1000) |
| Antibody | Rabbit monoclonal anti-caveolin-1 | CST | Cat. #: 3,267 | WB (1:1000) |
| Antibody | Rabbit monoclonal anti-phoso-AKT1 | CST | Cat. #: 5,012 | WB (1:1000) |
| Antibody | Rabbit monoclonal anti-phoso-AKT2 | CST | Cat. #: 8,599 | WB (1:1000) |
| Antibody | Rabbit polyclonal anti- phospho-HSL | Absin | Cat. #: abs139855 | WB (1:1000) |
| Antibody | PE Mouse Monoclonal Anti-rat CD31 | BD Pharmingen | Cat. #: 555,027RRID: AB_395657 | FACS(0.25 µg per million cells in 100 µl volume) |
| Antibody | FITC Mouse Monoclonal anti-rat CD45 | BioLegend | Cat. #: 202,205RRIS: AB_314005 | FACS(0.25 µg per million cells in 100 µl volume) |
| Antibody | PerCP/Cyanine5.5 Armenian Hamster Monoclonal Anti-mouse/rat CD29 | BioLegend | Cat. #: 102,227RRIS: AB_2572078 | FACS(0.25 µg per million cells in 100 µl volume) |
| Antibody | Pe-Cy7 Mouse Monoclonal Anti-mouse/rat/human CD34 | Santa Cruz Biotechnology | Cat. #: sc-7324RRIS: AB_2572078 | FACS(1 µg per million cells in 100 µl volume) |
| Peptide, recombinant protein | Recombinant human insulin | Sigma | Cat. #: 407,709 | |
| Commercial assay or kit | NEFA LabAssay | Wako Diagnostics | Cat. #: 294–63601 | |
| Chemical compound, drug | Liberase TL | Roche | Cat. #: 05401020001 | |
| Chemical compound, drug | Dexamethasone | Sigma | Cat. #: D4902 | |
| Chemical compound, drug | Indomethacin | Sigma | Cat. #: I8280 | |
| Chemical compound, drug | 3-isobutyl-1-methylxanthine (IBMX) | Sigma | Cat. #: I7018 | |
| Chemical compound, drug | BODIPY 493/503 | ThermoFisher | Cat. #: D3922 | |
| Software, algorithm | Aperio ImageScope | Leica |
Additional files
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Supplementary file 1
Weight loss in EGFRI-related clinical trials.
- https://cdn.elifesciences.org/articles/72443/elife-72443-supp1-v1.docx
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Supplementary file 2
Blood cell analysis of Vehicle and HFD rats.
WBC: white blood cells. W-SCR: WBC-small cell ratio. W-MCR: WBC-middle cell ratio. W-LCR: WBC-large cell ratio. W-SCC: WBC-small cell count. WBC-MCC: WBC-middle cell count. WBC-LCC: WBC-large cell count. RBC: red blood cells. HGB: Haemohlobin. HCT: Haematocrit. MCV: Mean corpuscular volume. MCH: Mean corpuscular haemoglobin. MCHC: Mean corpuscular haemoglobin concentration. RDW-SD: RBC-distribution width standard deviation. RDW-CV: RBC-distribution width variation coefficient. PLT: Platelets. PDW: Platelets distribution width. MPV: Mean platelet volume. P-LCR: Platelet large cell ratio. Data are presented as the means ± SEM. P < 0.05 using 2-tailed unpaired Student’s t test.
- https://cdn.elifesciences.org/articles/72443/elife-72443-supp2-v1.xlsx
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Supplementary file 3
Blood cell analysis of Vehicle and DCA rats.
WBC: white blood cells. W-SCR: WBC-small cell ratio. W-MCR: WBC-middle cell ratio. W-LCR: WBC-large cell ratio. W-SCC: WBC-small cell count. WBC-MCC: WBC-middle cell count. WBC-LCC: WBC-large cell count. RBC: red blood cells. HGB: Haemohlobin. HCT: Haematocrit. MCV: Mean corpuscular volume. MCH: Mean corpuscular haemoglobin. MCHC: Mean corpuscular haemoglobin concentration. RDW-SD: RBC-distribution width standard deviation. RDW-CV: RBC-distribution width variation coefficient. PLT: Platelets. PDW: Platelets distribution width. MPV: Mean platelet volume. P-LCR: Platelet large cell ratio. Data are presented as the means ± SEM. P < 0.05 using 2-tailed unpaired Student’s t test.
- https://cdn.elifesciences.org/articles/72443/elife-72443-supp3-v1.xlsx
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Supplementary file 4
Primer sequence information.
- https://cdn.elifesciences.org/articles/72443/elife-72443-supp4-v1.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/72443/elife-72443-transrepform1-v1.docx
