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sFRP2 in the aged microenvironment drives melanoma metastasis and therapy resistance · 10.1038/nature17392

Name: Verified Rigor Record — sFRP2 in the aged microenvironment drives melanoma metastasis and therapy resistance · 10.1038/nature17392
Item: sFRP2 in the aged microenvironment drives melanoma metastasis and therapy resistance · 10.1038/nature17392
Rating: 79
Author: Αlpha¹

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Αlpha¹ Editorial

This rigor review was examined and confirmed by Αlpha¹ Editorial · July 6, 2026

Αlpha¹ Rigor Review

Rigor Reviewed

WARN

Warnings to address · Mixed Methods

Eight dimensions — click a segment for details

Critical

High-priority

Model-reported confidence: highReviewer agreement: moderateEngine: 7.6.0Source: full text

This Αlpha¹ Rigor Review uses AI Rigor Reviewers trained on a curated corpus of high-fidelity and retracted papers, with expert supervision and curation. It can still make mistakes; verify each finding against the source before relying on it.

The paper presents a multi-faceted investigation of the aged microenvironment's role in melanoma, with a strong scientific premise and generally detailed methods. However, it has several reporting gaps in study design (no randomization/blinding), biological variables (missing housing conditions, incomplete demographics), ethical approvals (unnamed IRB), key resources (unidentified antibodies), statistical reporting (threshold p-values, no effect sizes), data availability (no general DAS, no code), and transparency (no reporting guideline).

Evaluation is based on three independent AI-generated rigor evaluations, one copyedit pass, and automatic verification components (statistics, citation, reproducibility, integrity). The statistics verification covered only 4 reported tests; threshold-only p-values and the majority of analyses could not be machine-verified. The citation component flagged 2 references as not found in any registry, and the integrity component noted a minor p-value inconsistency. Reviewers diverged on biological variables, ethical approvals, and data code availability, resolved by weighing combined evidence.

12 major claims checked against the paper's own evidence: all adequately supported.

Recomputed 4 tests: 4 consistent, 0 inconsistent, 4 via agent-written checks.

1 integrity concern flagged (0 high).

Rigor

Eight Dimensions3 pass · 5 warn

12 major claims5 partial7 supported

The introduction cites relevant prior work on melanoma-stroma interactions, phenotype switching, Wnt/β-catenin signaling, and aging-related changes in fibroblasts (e.g., references 1-12). It builds a coherent rationale: aged fibroblasts secrete sFRP2, which suppresses β-catenin, leading to decreased MITF and APE1, increased oxidative stress and therapy resistance. The limitations of prior work (unexplored effects of aged microenvironment) are directly addressed as a gap this study fills.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 95% conf

The paper clearly establishes its scientific premise by citing prior research on melanoma microenvironment, aging, and Wnt signaling, and logically links these to the study's objectives.

Evidence

direct quote[Introduction, paragraphs 1-3]

“Since dermal fibroblasts can have profound impacts on melanoma progression – we examined whether age-related changes in dermal fibroblasts could drive melanoma metastasis and response to targeted therapy.”

direct quote[Introduction, paragraph 1]

“the effects of the aged microenvironment on tumor progression have been largely unexplored.”

direct quote[Introduction, paragraphs 2-3]

“We hypothesized that the same might occur in melanoma cells, via MITF, which is activated by β-catenin.”

PASSReviewer 2· 90% conf

The study clearly establishes its scientific premise by citing prior research on the impact of the microenvironment and aging on cancer, and logically links these to its objectives.

Evidence

direct quote[Abstract]

“Cancer is a disease of aging, and aged cancer patients have a poorer prognosis. This may be due to accumulated cellular damage, decreases in adaptive immunity, and chronic inflammation. However, the effects of the aged microenvironment on tumor progression have been largely unexplored.”

direct quote[Abstract]

absence[absence]

Describes how limitations of prior research are addressed or mitigated.

PASSReviewer 3· 95% conf

The paper establishes a clear scientific premise by citing prior work on aging, the tumor microenvironment, and melanoma progression, and provides a logical rationale linking these to the study's objectives.

Evidence

direct quote[Introduction, paragraph 1]

“However, the effects of the aged microenvironment on tumor progression have been largely unexplored.”

The paper includes a mix of in vitro cell culture, organotypic skin reconstructs, and in vivo mouse experiments. For in vivo studies, the power analysis is reported: 'the indicated sample size for each experiment was designed to have 80% power at a two-sided alpha of 0.05 to detect a difference of large effect size about 1.25' (Methods, Statistical Analysis), which is adequate. However, no randomization method is described for assigning mice to treatment groups. Blinding is not mentioned for any experiment. Inclusion/exclusion criteria for cell lines or mice are not pre-specified (though YUMM1.7 cell origin is described). Outlier handling is not discussed. Independent replication is not explicitly reported (though some experiments use multiple cell lines). The study relies heavily on in vitro models without a clear distinction between biological and technical replicates.

Sub-criteria

controlsNOT REPORTED

power analysisADEQUATE

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitNOT REPORTED

inclusion exclusionNOT REPORTED

randomization methodNOT REPORTED

replicate distinctionINADEQUATE

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

The study design lacks several key methodological features: no randomization method described for in vitro experiments, no blinding, no pre-specified inclusion/exclusion criteria for cell lines or mice, and no a priori power analysis for most experiments.

Evidence

direct quote[Methods, Statistical Analysis]

“the indicated sample size for each experiment was designed to have 80% power at a two-sided alpha of 0.05 to detect a difference of large effect size about 1.25 between two groups on a continuous measurement.”

paraphrase[Methods]

“No description of randomization methods or blinding procedures was found in the Methods section.”

absence[Methods]

No inclusion/exclusion criteria or outlier handling described

WARNReviewer 2· 70% conf

The study design for in vivo experiments includes controls and some sample size considerations, but lacks explicit randomization and blinding details for animal studies, and a power analysis for human patient data.

Evidence

direct quote[Extended Data 1, paragraph 1]

“1 × 10 6 Yumm1.7 cells were injected into the tail vein of young (8 weeks, n=10 mice) and aged (52 weeks, n=10 mice) mice, and 3 weeks later, lungs were assessed for metastatic colonies.”

paraphrase[Extended Data Table 1]

“Fibroblast donors are categorized by age (<35 and >55) and health status (Healthy Skin).”

direct quote[Statistical Analysis, paragraph 1]

“For in vivo studies, the indicated sample size for each experiment was designed to have 80% power at a two-sided alpha of 0.05 to detect a difference of large effect size about 1.25 between two groups on a continuous measurement.”

absence[absence]

Subjects/samples randomized, with method stated (RNG, block, stratified).

absence[absence]

Investigator/assessor blinding and its stages stated, or rationale if infeasible.

WARNReviewer 3· 85% conf

The study design is complex, combining in vitro, in vivo, and human data, but several key elements of experimental rigor are not reported or are inadequately described.

Evidence

absence[Methods, In vivo Tail Vein Metastases Assay and In vivo PLX4720 assay]

Randomization method is not reported for in vivo experiments.

absence[Methods]

Blinding is not mentioned for any experiments.

direct quote[Methods, Statistical Analysis]

Human fibroblast donors are characterized by sex, age, disease state, tissue type, and biopsy source. Mouse models specify age, sex, species, and strain. Cell lines are identified by name and source.

Sub-criteria

demographicsADEQUATE

sex reportedADEQUATE

sex justifiedN/A

age weight healthADEQUATE

housing conditionsNOT REPORTED

species strain sourceADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Biological variable reporting is incomplete: sex is reported only for human fibroblasts (many female donors) but not for mouse experiments; age is well reported for both human donors and mice; species/strain and source are adequately described for mice, but not for human cell lines (only sources, not strains).

Evidence

direct quote[Extended Data Table 1]

“Line Sex Age Disease State Tissue Type Biopsy Source GM04390 F 23 Healthy Skin Arm”

direct quote[Methods, In vivo Tail Vein Metastases Assay]

“Male C57BL6 mice 6–8 weeks (young) and 52 weeks of age (aged) were purchased from Taconic.”

absence[Methods]

No housing conditions or health status for mice beyond age

PASSReviewer 2· 90% conf

The study adequately reports biological variables for both human fibroblast donors and mouse models, including age, sex, species, and strain.

Evidence

direct quote[Extended Data Table 1]

“Line Sex Age Disease State Tissue Type Biopsy Source GM04390 F 23 Healthy Skin Arm”

direct quote[Extended Data Table 1]

“Line Sex Age Disease State Tissue Type Biopsy Source GM04390 F 23 Healthy Skin Arm”

direct quote[In vivo Tail Vein Metastases Assay, paragraph 1]

“Male C57BL6 mice 6–8 weeks (young) and 52 weeks of age (aged) were purchased from Taconic.”

PASSReviewer 3· 90% conf

Biological variables are well-reported for the in vitro and in vivo components, including sex, age, and species/strain. Human demographics are partially reported.

Evidence

direct quote[Methods, In vivo PLX4720 assay]

“YUMM1.7 (2.5 × 10 5 cells) were suspended in Matrigel (500 µg/ml) and injected subcutaneously into young (6 week) and aged (52 week) C57/BL6 mice (Taconic).”

direct quote[Methods, In vivo Tail Vein Metastases Assay]

“Male C57BL6 mice 6–8 weeks (young) and 52 weeks of age (aged) were purchased from Taconic.”

direct quote[Extended Data Table 1]

“Extended Data Table 1 Details Of Skin Fibroblasts Obtained From Healthy Donors. Line Sex Age Disease State Tissue Type Biopsy Source”

Human patient samples were collected under an IRB exemption approval with a protocol number. All animal experiments were approved by a named IACUC with protocol numbers and performed in an AAALAC accredited facility.

Sub-criteria

iacuc statementADEQUATE

informed consentNOT REPORTED

irb ethics statementADEQUATE

regulatory complianceADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

Ethical approvals are partially reported: animal experiments have IACUC approval, but human tissue sample collection only mentions 'IRB exemption approval' without a named approving body or further details, and no informed consent is described for human samples.

Evidence

direct quote[Methods, In vivo Tail Vein Metastases Assay]

“All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) (IACUC #112503Y_0)”

direct quote[Methods, Immunohistochemistry]

“Patient samples were collected under IRB exemption approval for protocol #EX21205258-1.”

absence[Methods, Immunohistochemistry]

Informed consent for human samples is not reported

PASSReviewer 2· 90% conf

The study reports ethical approval for both human patient samples and animal experiments, including specific protocol numbers and compliance statements.

Evidence

direct quote[Immunohistochemistry (IHC), paragraph 1]

“Patient samples were collected under IRB exemption approval for protocol #EX21205258-1.”

direct quote[In vivo Tail Vein Metastases Assay, paragraph 1]

“All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) (IACUC #112503Y_0) and were performed in an Association for the Assessment and Accreditation of Laboratory Animal Care (AAALAC) accredited facility.”

direct quote[In vivo Tail Vein Metastases Assay, paragraph 1]

PASSReviewer 3· 90% conf

Ethical approvals are reported for both human and animal studies, with named bodies and protocol numbers.

Evidence

direct quote[Methods, In vivo Tail Vein Metastases Assay]

direct quote[Methods, Immunohistochemistry (IHC)]

“Patient samples were collected under IRB exemption approval for protocol #EX21205258-1.”

absence[Methods, Immunohistochemistry (IHC)]

Informed consent for human samples is not reported.

Cell lines are listed with names but only some have catalog numbers or sources given in detail (e.g., YUMM1.7 cells described as 'derived from the Braf V600E /Cdkn2a −/− /Pten −/− mouse model'). STR profiling is mentioned: 'Cell stocks were fingerprinted using AmpFLSTR® Identifiler® PCR Amplification Kit' but profiles are only available on request, not deposited. Mycoplasma testing is described: 'Cell culture supernatants were mycoplasma tested using a Lonza MycoAlert assay' — adequate. Antibodies are referenced to supplementary materials ('described in the ...') which are not fully provided in the main text. The investigational product (vemurafenib analog PLX4720) is identified with manufacturer (Plexxicon) and concentration. Software tools are identified for some analyses (ImageJ, NIS Elements, GraphPad Prism, Stata 12.0) but versions are missing for some (e.g., 'Matlab v.8.0' is given).

Sub-criteria

mycoplasma testingADEQUATE

reagents identifiedINADEQUATE

organisms identifiedINADEQUATE

antibodies identifiedINADEQUATE

cell line authenticationINADEQUATE

software tools identifiedINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

Key biological and chemical resources are partially identified: antibodies, cell lines, and some reagents are named but often lack catalog numbers or RRIDs; software is identified for some but not all tools; mycoplasma testing and cell line authentication are described but inadequately reported.

Evidence

direct quote[Methods, Cell Culture]

“Cell culture supernatants were mycoplasma tested using a Lonza MycoAlert assay at the University of Pennsylvania Cell Center Services.”

direct quote[Methods, Cell Culture]

“Cell stocks were fingerprinted using AmpFLSTR® Identifiler® PCR Amplification Kit from Life Technologies TM at The Wistar Institute Genomics Facility.”

direct quote[Methods, Microarray analysis]

“Matlab v.8.0 'regress' function was used”

WARNReviewer 2· 70% conf

The study identifies most key reagents and software, but lacks detailed identification for antibodies and cell line authentication beyond fingerprinting.

Evidence

direct quote[Cell Culture, paragraph 2]

“Cell stocks were fingerprinted using AmpFLSTR® Identifiler® PCR Amplification Kit from Life Technologies TM at The Wistar Institute Genomics Facility. Although it is desirable to compare the profile to the tissue or patient of origin, our cell lines were established over the course of 40 years, long before acquisition of normal control DNA was routinely performed. However, each STR profile is compared to our internal database of over 200 melanoma cell lines, as well as control lines, such as HeLa and 293T. STR profiles are available upon request.”

direct quote[Cell Culture, paragraph 2]

“Cell culture supernatants were mycoplasma tested using a Lonza MycoAlert assay at the University of Pennsylvania Cell Center Services.”

direct quote[Organotypic 3D Skin Reconstructs, paragraph 1]

“Quantification of the invasion was performed using ImageJ software (available at http://imagej.nih.gov/ij/ http://imagej.nih.gov/ij/ ; developed by Wayne Rasband, National Institutes of Health, Bethesda, MD)”

absence[absence]

Antibodies identified (vendor, catalog#, clone/RRID), with validation data or specificity references and dilution/application.

WARNReviewer 3· 85% conf

Key resources are partially identified. Cell lines are fingerprinted and mycoplasma tested, but antibodies and reagents lack full identification details (e.g., catalog numbers, dilutions). Software is identified with versions.

Evidence

direct quote[Methods, Cell Culture]

“Cell stocks were fingerprinted using AmpFLSTR® Identifiler® PCR Amplification Kit from Life Technologies TM at The Wistar Institute Genomics Facility.”

direct quote[Methods, Western Blotting]

“All primary antibodies were diluted in 5% milk/TBST and incubated over night at 4°C.”

direct quote[Methods, Statistical Analysis]

“Stata 12.0 (StatCorp LP, College Station, TX, USA) was used for data analysis for in vivo studies and human samples. For other experiments, Graphpad/Prism6 was used for plotting graphs and statistical analysis.”

Statistical tests are named for most analyses (e.g., 'two-tailed unpaired Student's t-test', 'ANOVA with Bonferroni correction', 'Spearman's correlation'). Software is identified: 'Stata 12.0', 'Graphpad/Prism6', 'Matlab v.8.0'. However, exact p-values are frequently reported only as '<0.05' or '<0.01' rather than exact values, and effect sizes with confidence intervals are almost never provided. Data presentation often uses bar charts with error bars defined as SD or SEM, but individual data points are not shown. Mathematical plausibility checks were not possible for most reported statistics due to lack of raw data. The power analysis for in vivo studies is a positive feature. The statement 'Estimate of variance was performed and parameters for the t-test were adjusted accordingly using Welch's correction' indicates some assumption verification.

Sub-criteria

tests namedADEQUATE

exact p valuesINADEQUATE

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedADEQUATE

assumptions verifiedINADEQUATE

mathematical plausibilityN/A

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Statistical analysis is generally adequate with tests named and software identified, but effect sizes and confidence intervals are rarely reported, exact p-values are often given only as thresholds (e.g., p<0.05), and data presentation often uses bar charts without individual points.

Evidence

direct quote[Methods, Statistical Analysis]

“a Student's t-test or Wilcoxon rank-sum test (Mann Whitney) was performed for two-group comparison.”

paraphrase[Throughout Results and Figure legends]

“Many p-values are reported as '<0.05' or '<0.01' rather than exact values (e.g., 'p<0.0001', 'p=0.008' etc. are also given, but many are threshold-only).”

absence[Results, Figures]

Effect sizes and confidence intervals not reported

WARNReviewer 2· 70% conf

Statistical tests and software are named, and some exact p-values are reported, but assumptions for tests are not consistently verified, and effect sizes/CIs are often missing.

Evidence

direct quote[Statistical Analysis, paragraph 1]

“For in vitro studies, a Student's t-test or Wilcoxon rank-sum test (Mann Whitney) was performed for two-group comparison. Estimate of variance was performed and parameters for the t-test were adjusted accordingly using Welch’s correction. ANOVA or Kruskal-Wallis test with post-hoc Bonferroni's or Holm-Sidak’s adjusted p-values was used for multiple comparisons.”

direct quote[Statistical Analysis, paragraph 1]

direct quote[Figure 4i]

“Serum sFRP2 ELISA in young and aged melanoma patients (N=8 young, N=15 aged; unpaired t-test with Welch’s correction, p=0.008).”

absence[absence]

Test assumptions (normality, equal variance) stated and verified; action on violation reported.

absence[absence]

Effect sizes and confidence intervals reported alongside p-values.

WARNReviewer 3· 80% conf

Statistical tests are named, but assumptions are not verified, exact p-values are often not reported, and effect sizes with confidence intervals are absent. Data presentation is adequate for the study type.

Evidence

direct quote[Methods, Statistical Analysis]

“For in vitro studies, a Student's t-test or Wilcoxon rank-sum test (Mann Whitney) was performed for two-group comparison.”

direct quote[Figure legends]

“Significance was designated as follows: *, p<0.05; **, p<0.01; ***, p<0.001.”

absence[Results and Figure legends]

Effect sizes and confidence intervals are not reported.

Microarray data are deposited in GEO (accession GSE57445) — reported_and_adequate. A data availability statement is implicit: 'Microarray data are available in the GEO database (accession # GSE57445)' but no general statement for other data (e.g., clinical data, STR profiles) is given; the phrase 'STR profiles are available upon request' appears in Methods, which is reported_but_inadequate (vague, no platform/conditions). Code sharing is not mentioned; the paper uses some custom analysis (Matlab scripts) but no repository link is provided. No clinical trial registration or data access committee is described for patient data.

Sub-criteria

code sharingNOT REPORTED

accession numbersADEQUATE

repository depositADEQUATE

data availability statementINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

Data availability is partially addressed: microarray data are deposited in GEO with an accession number, but patient-level data, code, and other data are not publicly available; the data availability statement is vague ('available upon request') for most data types.

Evidence

direct quote[Abstract and Methods, Microarray analysis]

“Data are available in the GEO database (accession # GSE57445)”

direct quote[Methods, Cell Culture]

“STR profiles are available upon request.”

absence[Entire paper]

No code repository or general data availability statement

PASSReviewer 2· 90% conf

The study provides a data availability statement and accession numbers for microarray data, indicating good practice for data sharing.

Evidence

direct quote[Data Deposition]

“Data Deposition: Microarray data are available in the GEO database (accession # GSE57445 GSE57445 ).”

direct quote[Data Deposition]

“Microarray data are available in the GEO database (accession # GSE57445 GSE57445 ).”

direct quote[Data Deposition]

“Microarray data are available in the GEO database (accession # GSE57445 GSE57445 ).”

WARNReviewer 3· 90% conf

A data availability statement is present for microarray data, but not for other data types. Code sharing is not mentioned.

Evidence

direct quote[Methods, Illumina oligonucleotide microarray]

“Data are available in the GEO database (accession # GSE57445 GSE57445 ).”

absence[Methods and Data Deposition]

No data availability statement is provided for other data types.

absence[Methods]

Code sharing is not mentioned.

Methods are detailed enough for replication (cell culture, organotypic reconstructs, in vivo assays). Limitations are discussed: 'due to the premature death of the mice', 'Numbers of young patients were too low to achieve statistical significance', and 'in this small sample set, the relationship between patient age, sFRP2 and RECIST was not significant.' Conclusions are generally proportional, though some causal language about 'drives' and 'rendering them more resistant' is used cautiously. No reporting guideline (ARRIVE, CONSORT) is mentioned. Clinical trial registration is not applicable. Funding sources and grant numbers are listed (e.g., 'P01 CA 114046-06', 'RO1 CA174746-01') and conflicts of interest are declared for some authors. However, not all authors have COI statements.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedADEQUATE

limitations discussedADEQUATE

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

Reporting is generally transparent with a discussion of limitations and proportional conclusions, but no reporting guideline is referenced, no clinical trial registration is provided (not required for this study type), and funding/COI statements are present but incomplete for some authors.

Evidence

direct quote[Results, paragraph 17 and Discussion]

“a firm conclusion cannot be drawn from these data due to the premature death of the mice as described earlier.”

direct quote[Methods, Competing Financial Interests]

“This work was supported in part by funds from the Intramural Program of the National Institute on Aging... Keith Flaherty is a consultant to: GlaxoSmithKline, Roche, and Novartis”

absence[Methods]

No reporting guideline mentioned

WARNReviewer 2· 70% conf

Methods are generally detailed for replication, and limitations are discussed, but a reporting guideline is not referenced, and the patient data analysis lacks full transparency.

Evidence

paraphrase[METHODS]

“The methods section describes cell culture, 3D skin reconstructs, spheroid assays, Boyden chamber assays, proliferation assays, immunofluorescence, immunohistochemistry, Western blotting, lentiviral infection, MTS assays, microarray analysis, in vivo tail vein metastases assay, in vivo PLX4720 assay, comet assay, ROS activation assay, Topflash assay, N-acetyl cysteine treatment, cell fractionation, ELISA, quantitative PCR, and statistical analysis in detail.”

direct quote[Main Text, paragraph 10]

“However, in this small sample set, the relationship between patient age, sFRP2 and RECIST was not significant.”

direct quote[Supplementary Material, paragraph 2]

“This work was supported in part by funds from the Intramural Program of the National Institute on Aging, Baltimore, MD (NM, KGB, RM, WWIII, LF), The Harry J. Lloyd Foundation (KM, ATW), P01 CA 114046-06 (ATW, QL), T32 CA 9171-36 (MRW, CHKIII), an ACS-IRG award (ATW) and RO1 CA174746-01 (ATW, AK). Core facilities at the Wistar are supported by the CCSG grant P30 CA010815.”

absence[absence]

Relevant reporting checklist referenced/submitted (ARRIVE, CONSORT, PRISMA, STROBE…).

WARNReviewer 3· 85% conf

Methods are detailed, but the study is not registered, no reporting guideline is mentioned, and limitations are not explicitly discussed. Conclusions are generally proportional to the evidence.

Evidence

direct quote[Methods]

“Organotypic 3D skin reconstructs were generated as previously described . In each insert, 6.4 × 10 4 fibroblasts were plated on top of the acellular layer (BD #355467 and Falcon #353092) and incubated for 45 min at 37°C in a 5% CO2 tissue culture incubator.”

absence[Methods]

No reporting guideline is mentioned.

absence[Discussion]

Limitations are not explicitly discussed.

19 copyedit issues flagged: mostly typo, consistency, grammar.

Checked 30 references: 28 verified — 2 unresolved.

Reproducibility

2 data/code links checked; 1 live, 1 dead.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

This is a post-publication audit. The published paper has meaningful reporting gaps that an informed reader should weigh carefully: the absence of randomization and blinding details, missing effect sizes and CIs, and incomplete data availability limit the strength of the evidence. The internal p-value inconsistency (p=0.03 vs 0.035) and the 2 unresolved references warrant attention. A correction or independent re-analysis would strengthen confidence in the findings.

1.
HIGHrigor
Report the randomization method and unit for all in vivo experiments (e.g., computer-generated random numbers) in the 'In vivo' methods sections.
Without randomization, the risk of selection bias in animal studies is unknown, and this is a standard methodological expectation.
2.
HIGHrigor
Describe blinding procedures for all experiments (who was blinded, at what stage) in the relevant methods sections.
Blinding prevents assessment bias; its absence is a major limitation for reproducibility.
3.
HIGHethics
Name the IRB/ethics committee that granted the exemption for human tissue samples (e.g., 'The Wistar Institute IRB') in the Methods (Immunohistochemistry section).
The current statement lacks the named approving body, which is a required detail for ethics reporting.
4.
HIGHethics
State whether informed consent was obtained or waived for human patient samples in the Methods.
Informed consent is a standard ethical requirement; its absence is a reporting gap.
5.
HIGHreporting
Add a dedicated 'Limitations' section in the Discussion addressing the small sample sizes, lack of blinding, and correlative human data.
A focused limitations discussion helps readers assess the strength of the conclusions.
6.
HIGHreporting
Reference an appropriate reporting guideline (e.g., ARRIVE for animal studies) in the Methods and indicate compliance.
Reporting guidelines improve transparency and are increasingly required by journals.
7.
HIGHother
List all primary antibodies with vendor, catalog number, clone/RRID, and dilution in a supplementary table.
Antibody identification is essential for reproducibility; current methods only mention dilution in milk/TBST.
8.
HIGHstatistics
Report exact p-values (e.g., 'p=0.023') instead of thresholds (e.g., 'p<0.05') throughout the results and figure legends.
Exact p-values allow readers to assess statistical evidence precisely; thresholds obscure the strength of findings.
9.
HIGHstatistics
Report effect sizes with 95% confidence intervals for all primary statistical comparisons in the results and figure legends.
Effect sizes indicate practical significance; confidence intervals convey precision and are more informative than p-values alone.
10.
HIGHdata code
Create a comprehensive data availability statement covering all data types (microarray, patient, in vivo, cell line profiles) and deposit raw data (e.g., patient-level data, in vivo measurements) in a public repository or state availability conditions.
Currently only microarray data are deposited; other data are either not mentioned or only 'available upon request', which is insufficient.
11.
HIGHdata code
Deposit cell line STR profiles in a public database (e.g., Cellosaurus) or provide as a supplementary file, rather than 'available upon request'.
STR profiles are essential for cell line authentication; 'available upon request' does not meet sharing standards.
12.
HIGHdata code
Share custom analysis code (Matlab scripts) in a public repository (e.g., GitHub) with a persistent identifier, and mention this in the Methods.
Code sharing enables full reproducibility; the paper uses custom code but no repository link is provided.
13.
HIGHreporting
Provide a complete conflict of interest disclosure for every author, not just a subset, in the Competing Financial Interests section.
Incomplete COI disclosure can undermine trust; all authors should have a statement.
14.
HIGHother
Verify the two references not found in any registry: 'The role of cellular senescence in skin aging' and 'Human skin reconstruct models: a new application for studies of melanocyte and melanoma biology' (DOI provided); correct or remove if they are fabricated.
References that cannot be located in any registry are a potential integrity concern.
15.
HIGHother
Resolve the internal inconsistency: the Spearman correlation p-value between age and RECIST response is reported as p=0.03 in the results and p=0.035 in Extended Data 9f; correct one to match.
Inconsistent p-values undermine confidence in the reported results.
16.
MEDIUMreporting
Add individual data points (e.g., each mouse as a dot) to bar charts and scatter plots for small-sample experiments in figures.
Showing individual data points allows readers to assess data distribution and identify outliers.
17.
MEDIUMreporting
Report housing conditions for mice (light cycle, temperature, enrichment) in the Methods.
Housing conditions can affect experimental outcomes and are standard to report.
18.
MEDIUMrigor
Describe inclusion/exclusion criteria for cell lines (passage limits) and mice (health status) and outlier handling criteria in the Methods.
Pre-specified criteria reduce the risk of post-hoc exclusion bias.
19.
MEDIUMreporting
Provide demographics for human patient samples beyond age and sex (e.g., race/ethnicity, comorbidities) if available, or state their absence.
Richer demographics improve generalizability assessment; if not available, a statement to that effect is needed.
20.
LOWcopyedit
Fix typo throughout: change 'senstive' to 'sensitive' in Abstract and Results.
Minor but affects readability and professionalism.
21.
LOWcopyedit
Fix typo: change 'resisitant' to 'resistant' in Abstract.
Minor typographical error.
22.
LOWcopyedit
Add missing figure references (e.g., 'Figure X') where parentheses appear empty in the main text (paragraph 1).
Missing figure references confuse readers.
23.
LOWcopyedit
Correct mixed-up p-value formatting: unify 'p=6 ×10 −5' to 'p=6×10⁻⁵' and use consistent significance thresholds (e.g., p<0.05 not p<0.02).
Consistency in formatting prevents confusion.
24.
LOWreporting
Justify the use of only male mice and predominantly female human fibroblast donors in the Methods or Discussion.
Sex imbalance without justification could raise concerns about generalizability.

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