Vascularized human skin assembloids model compartmental skin organization and immune responsiveness in vitro

May combine two claims.

SkOs often display inverted polarity, with the epidermis positioned internally and the dermis externally, and frequently undergo ectopic cartilage differentiation.

“They often display inverted polarity, in which the epidermis is positioned internally and the dermis externally, and frequently undergo ectopic cartilage differentiation 14,15 .”

• cites [14] Lee, J. et al. Hair-bearing human skin generated entirely from pluripotent stem cells. Nature 582, 399–404 (2020). sourcePartly supported

  The passage on organoid induction (page unavailable) describes an inner epithelial layer surrounded by outer mesenchymal cells, supporting the polarity description. The passage on late-stage organoids reports cartilage developing in the tail region, but does not establish how frequently it occurs or describe it as ectopic; it instead relates cartilage to the organoids’ craniofacial tissue architecture. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites [15] Mostina, M. et al. Coordinated development of immune cell populations in vascularized skin organoids from human induced pluripotent stem cells. Adv. Healthc. Mater. 14, e02108 (2025). sourcePartly supported

  The Results section, “Endothelialized Skin Organoids Recapitulate Endothelial Structures in Vitro” (page unavailable), reports abundant cartilage in the SKO + GFP-ECFC group, supporting cartilage formation in that condition. It does not establish that cartilage differentiation is frequent across SKOs. The Results describe epithelial cells shedding into an organoid cyst but do not state that SKOs often have an internal epidermis and external dermis. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

hiPSC-derived blood vessel organoids yield self-assembled endothelial networks with lumina and mural support cells capable of forming perfusable vascular structures.

“In parallel, advances in the generation of hiPSCderived blood vessel organoids (BVOs) have yielded selfassembled endothelial networks with lumina and mural support cells capable of forming perfusable vascular structures.”

• cites [15] Mostina, M. et al. Coordinated development of immune cell populations in vascularized skin organoids from human induced pluripotent stem cells. Adv. Healthc. Mater. 14, e02108 (2025). sourcePartly supported

  The Results section, “Human Vascular Organoids Recapitulate Broad Blood Vessel Phenotypes,” reports hiPSC-derived vascular organoids with endothelial cells, pericytes, mural cells, and capillary structures. The Results section on vascularized skin organoids also describes endothelial capillaries supported by pericytes. Our copy does not establish that the networks have lumina or can form perfusable structures. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [17] Wimmer, R. A. et al. Human blood vessel organoids as a model of diabetic vasculopathy. Nature 565, 505–510 (2019). sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Transient activation of WNT signalling during organoid development suppresses ectopic cartilage formation and promotes generation of large, correctly oriented skin organoids suitable for modelling solar UV-induced damage and repair.

“Transient activation of WNT signalling during organoid development suppresses ectopic cartilage formation and promotes the generation of large, correctly oriented skin organoids suitable for modelling solar UV-induced damage and repair 16 .”

• cites [16] Kim, M.-J. et al. Modeling of solar UV-induced photodamage on the hair follicles in human skin organoids. J. Tissue Eng. 15, 20417314241248753 (2024). sourcePartly supported

  The Methods (page unavailable) say a WNT agonist was added during organoid development to prevent off-target cartilage differentiation and increase organoid diameter. The Discussion (page unavailable) attributes the planar, skin-like arrangement to cutting the organoids and using an air–liquid interface, not to WNT activation alone. The paper uses those organoids to model solar UV damage and a potential treatment, but does not establish that WNT activation itself produces their orientation. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim. May combine two claims.

Skin organoids derived from hiPSCs recapitulate epidermal and dermal differentiation and generate appendages such as hair follicles, melanocytes and neurons.

“Skin organoids (SkOs) derived from hiPSCs recapitulate epidermal and dermal differentiation and generate appendages such as hair follicles, melanocytes and neurons 13,14 .”

• cites [13] Jung, S.-Y. et al. Wnt-activating human skin organoid model of atopic dermatitis induced by Staphylococcus aureus and its protective effects by Cutibacterium acnes. Iscience 25, 105150 (2022). sourceSupported
• cites [14] Lee, J. et al. Hair-bearing human skin generated entirely from pluripotent stem cells. Nature 582, 399–404 (2020). sourceSupported

Classified as probably not a substantial claim. May combine two claims.

Assembloids have been applied to model complex inter-organ interactions, including vascularized tumour systems and brain-immune interfaces, enabling functional readouts such as immune infiltration and therapeutic response.

“Assembloids have recently been applied to model complex inter-organ interactions, including vascularized tumour systems and brain-immune interfaces, thereby enabling functional readouts such as immune infiltration and therapeutic response 19,20 .”

• cites [19] Zhou, R. et al. Vascularised organoids: recent advances and applications in cancer research. Clin. Transl. Med. 15, e70258 (2025). sourcePartly supported

  The ‘Drug responsiveness in tumour organoids’ section (page unavailable) describes coupled liver–breast cancer models and therapeutic-response readouts, supporting the inter-organ and vascularised-tumour parts of the claim. The copy does not describe brain–immune interfaces or immune-infiltration readouts, and ‘assembloids’ appears only in a reference title; it does not establish that the discussed models are assembloids. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [20] Kong, D. et al. CD19 CAR-expressing iPSC-derived NK cells effectively enhance migration and cytotoxicity into glioblastoma by targeting to the pericytes in tumor microenvironment. Biomed. Pharmacother. 174, 116436 (2024). sourceNot checked — no full text of the source

May combine two claims.

JAK1 inhibition and IL-4Rα antagonism both lead to suppression of STAT6 activation, thereby reducing downstream type 2-associated inflammatory signalling within the HDM-VSA model.

“These results suggest that JAK1 inhibition and IL-4Rα antagonism both lead to suppression of STAT6 activation 34,35 , thereby reducing downstream type 2-associated inflammatory signalling within the HDM-VSA model.”

• cites [34] Keegan, A. D., Leonard, W. J. & Zhu, J. Recent advances in understanding the role of IL-4 signaling. Fac. Rev. 10, 71 (2021). sourceSupported
• cites [35] Sharma, M. et al. Human germline heterozygous gain-of-function STAT6 variants cause severe allergic disease. J. Exp. Med. 220, e20221755 (2023). sourcePartly supported

  The Results section (‘JAK inhibitors and IL-4Rα monoclonal antibody can be used as potential therapeutics’; page unavailable) reports that ruxolitinib and tofacitinib reduced STAT6 activation and that dupilumab inhibited its IL-4-mediated increase. The paper calls the drugs JAK inhibitors but does not establish that JAK1 inhibition specifically produced the effect. The HDM-VSA interpretation follows the citations in the citing sentence and is not attributed to this source. (LLM, conf=0.82)

  Experimental — accuracy not yet measured

May combine two claims.

Reconstructed skin equivalents and organoid models lack integrated vascular and immune-associated compartments, limiting their capacity to model dynamic inflammatory processes.

“Although reconstructed skin equivalents and organoid models have advanced our understanding of epidermal differentiation, they lack integrated vascular and immune-associated compartments, limiting their capacity to model dynamic inflammatory processes 38,39 .”

• cites [38] Vilela de Sousa, I. et al. Skin models of cutaneous toxicity, transdermal transport and wound repair. Burns Trauma 11, tkad014 (2023). sourcePartly supported

  The Review’s “Microfluidic strategies for skin emulation” section reports that adding microvasculature to a skin-on-a-chip enabled cytokine production and leukocyte infiltration, supporting the importance of vascular and immune-related features for modelling inflammation. It does not establish that reconstructed skin equivalents and organoid models generally lack both compartments or that this specifically limits all such models. Page numbers were not available in our copy. (LLM, conf=0.80)

  Experimental — accuracy not yet measured
• cites [39] Quílez, C. et al. Targeting the complexity of in vitro skin models: a review of cutting-edge developments. J. Invest. Dermatol. 144, 2650–2670 (2024). sourcePartly supported

  The review says simple 3D skin models lack circulatory flow (New Advances, p. 4) and that cytokine-based models may not capture the dynamics of inflammation in vivo (Inflammatory skin diseases models, p. 11). But it also describes models incorporating immune cells and skin-on-chip models with vascularization (pp. 6, 11). The claim overgeneralizes these limitations to reconstructed skin equivalents and organoids as a whole. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Dupilumab is an IL-4Rα antagonist that blocks IL-4/IL-13 signalling.

“Upadacitinib, a JAK inhibitor with proven efficacy in moderate-tosevere AD 30,31 , and dupilumab, an IL-4Rα antagonist that blocks IL-4/IL-13 signalling 32,33 , were selected.”

• cites [32] Le Floc’h, A. et al. Dual blockade of IL-4 and IL-13 with dupilumab, an IL-4Rα antibody, is required to broadly inhibit type 2 inflammation. Allergy 75, 1188–1204 (2020). sourceSupported
• cites [33] Simpson, E. L. et al. Two phase 3 trials of dupilumab versus placebo in atopic dermatitis. N. Engl. J. Med. 375, 2335–2348 (2016). sourceSupported

May combine two claims.

Previous studies have shown that skin organoids can generate stratified epidermal and dermal compartments, but they generally fail to establish vascular structures, a defining feature of native human skin.

“Previous studies have shown that skin organoids can generate stratified epidermal and dermal compartments, but they generally fail to establish vascular structures, a defining feature of native human skin 13,16 .”

• cites [13] Jung, S.-Y. et al. Wnt-activating human skin organoid model of atopic dermatitis induced by Staphylococcus aureus and its protective effects by Cutibacterium acnes. Iscience 25, 105150 (2022). sourcePartly supported

  The Discussion reports that this paper’s ALI-skin organoids have a dermis beneath a stratified epidermis, and the Limitations of the study section explicitly says the system lacks vasculature (page numbers unavailable). Those passages support the claim for this model, but do not establish that skin organoids generally fail to form vascular structures. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [16] Kim, M.-J. et al. Modeling of solar UV-induced photodamage on the hair follicles in human skin organoids. J. Tissue Eng. 15, 20417314241248753 (2024). sourcePartly supported

  The Results (Characterization of iPSC-derived hair-bearing SkOs; page unavailable) describe organoids with a stratified epidermis and dermis. The paper does not state that skin organoids generally fail to form vascular structures; searches for vascular structures and vascularization found no supporting passage. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

May combine two claims.

HDM extract is a clinically relevant allergen that triggers type 2-associated inflammatory responses in human skin.

“To further evaluate its disease relevance, VSAs were challenged with HDM extract, a clinically relevant allergen that triggers type 2-associated inflammatory responses in human skin 44 .”

• cites [44] Humeau, M., Boniface, K. & Bodet, C. Cytokine-mediated crosstalk between keratinocytes and T cells in atopic dermatitis. Front. Immunol. 13, 801579 (2022). sourcePartly supported

  The Introduction (page unavailable) describes environmental allergens as factors in atopic dermatitis and identifies a strong type 2 response in affected skin. It does not identify house dust mite (HDM) extract or say that HDM extract triggers that response in human skin; searches for HDM, house dust mite, and Dermatophagoides did not find such a passage. (LLM, conf=0.83)

  Experimental — accuracy not yet measured

May combine two claims.

The current model does not yet capture inter-patient variability in pharmacological responses or the full chronicity of clinical AD.

“These pharmacological responses are mechanistically consistent with the known mechanisms of these therapeutics, whereas the current model does not yet capture inter-patient variability in pharmacological responses or the full chronicity of clinical AD [46][47][48] .”

• cites [46] Hamilton, J. D. et al. Dupilumab suppresses type 2 inflammatory biomarkers across multiple atopic, allergic diseases. Clin. Exp. Allergy 51, 915–931 (2021). sourceCould not verify

  The Methods (Studies and patients) describe clinical dupilumab trials, and the Results (Serum TARC) mention a statistical ANCOVA model. Neither passage addresses whether the citing authors’ current model captures inter-patient variability in drug responses or the chronicity of clinical AD. Searches for those model limitations found no relevant passage in our copy; page numbers were not provided. (LLM, conf=0.75)

  Experimental — accuracy not yet measured
• cites [47] Rodrigues, M. A. & Torres, T. JAK/STAT inhibitors for the treatment of atopic dermatitis. J. Dermatolog. Treat. 31, 33–40 (2020). sourceNot checked — no full text of the source
• cites [48] Blauvelt, A. et al. Efficacy and safety of upadacitinib vs dupilumab in adults with moderate-to-severe atopic dermatitis: a randomized clinical trial. JAMA Dermatol. 157, 1047–1055 (2021). sourceCould not verify

  The Discussion > Limitations section says the trial reports outcomes through 24 weeks and that longer-term data are being collected. It does not discuss the citing authors’ model or whether that model captures inter-patient variability or the chronicity of clinical AD. Searches for those model limitations found no supporting passage in our copy. (LLM, conf=0.77)

  Experimental — accuracy not yet measured

May combine two claims.

At D7, SkOs fused with the prevascularized ECM and spread along its surface, initiating structural integration.

“At D7, SkOs fused with the prevascularized ECM and spread along its surface, initiating structural integration.”

May combine two claims.

VSAs support immune co-culture and provide an experimentally accessible architecture for epidermal-side allergen exposure.

“Unlike conventional organoid models, VSAs supported immune co-culture and provided an experimentally accessible architecture for epidermal-side allergen exposure, enabling analysis of vascular-associated inflammatory responses and pharmacological modulation.”

May combine two claims.

Treatment of VSAs with upadacitinib and dupilumab improves barrier-associated features and suppresses STAT6-associated inflammatory signalling.

“Treatment with clinically used atopic dermatitis therapeutics upadacitinib (JAK1 inhibitor) and dupilumab (IL-4Rα antagonist) improves barrier-associated features and suppresses STAT6associated inflammatory signalling, showing responses consistent with their known mechanisms of action.”

May combine two claims.

VSAs display compartmental organization comprising a stratified epidermis, a fibroblast-rich dermal compartment, hair follicle- and neural-associated elements, and interconnected endothelial networks with luminal structures.

“VSAs displayed compartmental organization comprising a stratified epidermis, a fibroblast-rich dermal compartment, hair follicle-and neural-associated elements, and interconnected endothelial networks with luminal structures.”

May combine two claims.

At D0, SkOs and BVOs were physically juxtaposed within the matrix but retained distinct boundaries.

“At D0, SkOs and BVOs were physically juxtaposed within the matrix but retained distinct boundaries.”

May combine two claims.

At D15, the fused structure stabilized and increased in thickness, consistent with early tissue consolidation.

“At D15, the fused structure stabilized and increased in thickness, consistent with early tissue consolidation.”

May combine two claims.

VSAs exhibited elevated nitric oxide production and increased VEGF secretion compared with non-vascularized controls, indicating endothelial-associated activity.

“Functionally, VSAs exhibited elevated nitric oxide production and increased VEGF secretion compared with non-vascularized controls, indicating endothelialassociated activity (Fig. 1h, i).”

May combine two claims.

Immune cells were largely absent from the upper dermis in controls, whereas in HDM-treated VSAs they were observed along vascular structures within the upper dermal regions.

“Immune cells were largely absent from the upper dermis in controls, whereas in HDM-treated VSAs they were observed along vascular structures within the upper dermal regions (Fig. 6c, Supplementary Fig. 5c,d).”

May combine two claims.

Immunofluorescence analysis confirmed the presence of CD31+ vascular structures in both groups and demonstrated robust ICAM1 upregulation in LPS-treated VSAs, localized primarily to CD31+ vascular structures, with infiltrating MNCs situated in close proximity to ICAM1+ endothelial segments.

“Consistent with these findings, immunofluorescence analysis confirmed the presence of CD31 + vascular structures in both groups and demonstrated robust ICAM1 upregulation in LPS-treated VSAs, localized primarily to CD31 + vascular structures, with infiltrating MNCs situated in close proximity to ICAM1 + endothelial segments (Fig. 4e, f).”

May combine two claims.

Human CD31+ vascular structures were detected within the grafted region and were closely associated with mouse CD31+ host vasculature.

“Human CD31 + vascular structures were detected within the grafted region and were closely associated with mouse CD31 + host vasculature (Supplementary Fig. 3h).”

May combine two claims.

Dupilumab increased FLG expression and reduced TARC/TSLP levels, with responses approaching saturation above 10 μg ml-1 without further benefit.

“Dupilumab also increased FLG expression and reduced TARC/TSLP levels, with responses approaching saturation above 10 μg ml -1 without further benefit (Supplementary Fig. 6b).”

May combine two claims.

Expression of STAT6-associated target genes, including TARC, MDC, CCL26 and POSTN, was strongly induced by HDM but attenuated by both agents.

“Expression of STAT6-associated target genes, including TARC, MDC, CCL26 and POSTN, was strongly induced by HDM but attenuated by both agents (Fig. 8g).”

May combine two claims.

Single-cell transcriptomic analyses show that VSAs retain diverse skin-associated lineages and exhibit increased endothelial and perivascular marker-defined populations.

“Together, these single-cell transcriptomic analyses show that VSAs retain diverse skin-associated lineages and exhibit increased endothelial and perivascular marker-defined populations, consistent with the vascular structures observed by histological and immunofluorescence analyses.”

May combine two claims.

Upadacitinib or dupilumab significantly reduced HDM-induced epidermal hyperplasia, shifting epidermal thickness towards control levels.

“HDM-treated VSAs exhibited pronounced epidermal hyperplasia, which was significantly reduced by upadacitinib or dupilumab, shifting epidermal thickness towards control levels (Fig. 7b, c).”

May combine two claims.

Upadacitinib increased FLG expression in a concentration-dependent manner up to 1 μM, whereas higher concentrations reduced efficacy and induced rebound increases in TARC/TSLP.

“Upadacitinib increased FLG expression in a concentration-dependent manner up to 1 μM, whereas higher concentrations reduced efficacy and induced rebound increases in TARC/TSLP (Supplementary Fig. 6a).”

May combine two claims.

Upon LPS stimulation, MNCs accumulated within the upper dermal region of VSAs and were frequently observed in close proximity to CD31+ vascular structures, whereas in matched nonvascularized SkOs, infiltrating cells rarely progressed beyond the lower dermis.

“Upon LPS stimulation, MNCs accumulated within the upper dermal region of VSAs and were frequently observed in close proximity to CD31 + vascular structures, whereas in matched nonvascularized SkOs, infiltrating cells rarely progressed beyond the lower dermis (Fig. 4b).”

May combine two claims.

Within the CD45+CD3- population, CD11c+HLA-DR+ cells were detected in both control and HDM-treated VSAs and were modestly increased following HDM stimulation.

“Within the CD45 + CD3 -population, CD11c + HLA-DR + cells were detected in both control and HDM-treated VSAs and were modestly increased following HDM stimulation (Fig. 6e; Supplementary Fig. 5f).”

May combine two claims.

CD11c+CD1c+ populations were identified within the CD45+CD3- compartment, supporting the presence of myeloid/APC-like immune populations in the VSA-MNC co-culture system.

“In addition, CD11c + CD1c + populations were identified within the CD45 + CD3 -compartment, supporting the presence of myeloid/APC-like immune populations in the VSA-MNC co-culture system (Supplementary Fig. 5g).”

May combine two claims.

CD31+ endothelial networks in VSAs were positioned beneath the KRT14+ basal epidermis, ensheathed by α-SMA+ perivascular cells and surrounded by Collagen IV deposition aligned with VE-cadherin+ junctions, forming vessel-like tubes associated with basement-membrane components.

“CD31 + endothelial networks were positioned beneath the KRT14 + basal epidermis, ensheathed by α-SMA + perivascular cells and surrounded by Collagen IV deposition aligned with VE-cadherin + junctions, together forming vessel-like tubes associated with basement-membrane components (Fig. 3h).”

May combine two claims.

Both SkOs and VSAs developed a stratified epidermis above a fibroblast-rich dermis.

“Histological analysis revealed that both models developed a stratified epidermis above a fibroblast-rich dermis; however, VSAs uniquely exhibited an expanded dermal region containing lumenized vascular structures, showing structural features reminiscent of adult skin (Fig. 3a).”

May combine two claims.

HDM-treated VSAs displayed AD-associated gene-expression patterns that partially paralleled those observed in skin from patients with AD, including type 2/alarmin-associated genes (IL33, IL4R, IL13RA2, CCL17, CCL26 and GATA3), keratinocyte activation and stress-response genes (POSTN, KRT16, S100A8, S100A9, PI3 and SERPINB4), and adhesion/infiltration-associated mediators (CXCL8, CXCL10, ICAM1 and

“A z-score- normalized heatmap showed that HDM-treated VSAs displayed AD-associated gene-expression patterns that partially paralleled those observed in skin from patients with AD, including type 2/ alarmin-associated genes (IL33, IL4R, IL13RA2, CCL17, CCL26 and GATA3), keratinocyte activation and stress-response genes (POSTN, KRT16, S100A8, S100A9, PI3 and SERPINB4), and adhesion/infiltration-ass”

May combine two claims.

Transcriptomic data from upadacitinib-treated VSAs showed concurrent downregulation of type 2-associated cytokine genes and improvement of barrier-related programmes.

“Transcriptomic data from upadacitinib-treated VSAs supported these findings, showing concurrent downregulation of type 2-associated cytokine genes and improvement of barrier-related programmes (Supplementary Fig. 7).”

May combine two claims.

Following LPS stimulation, avascular SkOs showed only modest induction, whereas vascularized constructs exhibited markedly higher responses, with the strongest activation observed in VSA+MNCs.

“Following LPS stimulation, avascular SkOs showed only modest induction, whereas vascularized constructs exhibited markedly higher responses, with the strongest activation observed in VSA+MNCs.”

May combine two claims.

Gene ontology analysis highlighted biological processes related to inflammatory response, cornification, immune activation and type 2 immune regulation, whereas KEGG enrichment identified cytokine-cytokine receptor interaction, TNF signalling and IL-17 signalling cascades.

“Gene ontology analysis highlighted biological processes related to inflammatory response, cornification, immune activation and type 2 immune regulation, whereas Kyoto Encyclopedia of Genes and Genomes enrichment identified cytokine-cytokine receptor interaction, TNF signalling and IL-17 signalling cascades, pathways implicated in AD-associated inflammation (Fig. 5c, d).”

May combine two claims.

VSAs treated with Up or Du displayed markedly reduced infiltration in the upper dermis, with immune cells largely confined to the lower dermis.

“By contrast, VSAs treated with Up or Du displayed markedly reduced infiltration in this region, with immune cells largely confined to the lower dermis (Fig. 8a, b).”

May combine two claims.

VSA grafts persisted in BALB/c nude mice after 4 weeks and contained human nuclear antigen-positive graft-derived tissue.

“Finally, in vivo transplantation analysis showed that VSA grafts persisted in BALB/c nude mice after 4 weeks and contained human nuclear antigen-positive graft-derived tissue (Supplementary Fig. 3f, g).”

May combine two claims.

CD31 and VE-cadherin (CD144) formed continuous, network-like structures resembling vessel junctions in VSAs, whereas SkOs lacked such organization.

“CD31 and VE-cadherin (CD144) formed continuous, network-like structures resembling vessel junctions in VSAs, whereas SkOs lacked such organization (Fig. 3g).”

May combine two claims.

Transcriptomic profiling revealed extensive reprogramming in HDM-treated VSAs, with hierarchical clustering clearly separating control and HDM groups.

“Transcriptomic profiling revealed extensive reprogramming in HDM-treated VSAs, with hierarchical clustering clearly separating control and HDM groups, indicative of a reproducible transcriptional divergence (Fig. 5b).”

May combine two claims.

VSAs contain organized vessel-like networks, marked by CD31 and CD144 and perivascular markers α-SMA and COL4, associated with the basement membrane and perivascular compartment.

“Immunostaining for vascular endothelial markers CD31 and CD144 (red/green) and perivascular markers α-SMA, COL4 and CD144 showing organized vessel-like networks associated with the basement membrane and perivascular compartment.”

May combine two claims.

HDM-induced STAT6 phosphorylation in whole-VSA lysates is reduced by upadacitinib and dupilumab, with total STAT6 unchanged.

“Western blot analysis and quantification of p-STAT6/STAT6 ratios using whole-VSA lysates, showing that HDM-induced STAT6 phosphorylation was reduced by Up and Du, with total STAT6 unchanged.”

May combine two claims.

After upadacitinib or dupilumab treatment, FLG and CLDN1 expression is increased and KRT16 expression is suppressed in HDM-treated VSAs.

“Immunofluorescence staining and quantification showing increased filaggrin (FLG) (red) and CLDN1 (green) expression and suppression of KRT16 (red) after Up or Du treatment.”

May combine two claims.

In vascularized skin assembloids (VSAs), SkOs positioned on pre-vascularized extracellular matrix expanded by D7, stabilized in thickness by D15, and exhibited extensive vascular branching by D28.

“SkOs were positioned on pre-vascularized extracellular matrix at D0, expanded by D7, stabilized in thickness by D15 and exhibited extensive vascular branching by D28 (arrowheads).”

May combine two claims.

CD4+ T cells accumulate in the upper dermis of HDM-treated VSAs, and this accumulation is reduced after upadacitinib or dupilumab treatment.

“Representative immunofluorescence images of CD4 + T cells (green) showing accumulation in the upper dermis of house dust mite (HDM)-treated vascularized skin assembloids (VSAs) and reduced accumulation after upadacitinib (Up) or dupilumab (Du) treatment.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Sample size represents the number of biologically independent replicates, with at least three independent experiments performed for each condition.

“Sample size (n) represents the number of biologically independent replicates, with at least three independent experiments performed for each condition.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Animal experiments were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee of Seoul National University (IACUC No. SNU-250310-3-1).

“All animal experiments were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee of Seoul National University (IACUC No. SNU-250310-3-1).”

Probably not a substantial claim: may not pass S3 (load-bearing).

For single-cell analyses, proportions of marker-positive cells were compared using Fisher's exact test with Benjamini-Hochberg correction for multiple testing.

“For single-cell analyses, proportions of marker-positive cells were compared using Fisher's exact test with Benjamini-Hochberg correction for multiple testing.”

Probably not a substantial claim: may not pass S3 (load-bearing).

This study was carried out in accordance with the approved guidelines of the Seoul National University Institutional Review Board (IRB-No.2504/003-001).

“This study was carried out in accordance with the approved guidelines of the Seoul National University Institutional Review Board (IRB-No.2504/003-001).”

May combine two claims.

This assembly approach supports coordinated epidermal differentiation, dermal-vascular organization and endothelial activity.

“This assembly approach supports coordinated epidermal differentiation, dermal-vascular organization and endothelial activity, thereby providing a foundation for constructing human-relevant VSA models.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Umbilical cord blood was obtained with the approval of the Seoul National University Institutional Review Board (IRB-No.E2405/002-021).

“Umbilical cord blood (UCB) was obtained from the Seoul metropolitan Government with the approval of the Seoul National University Institutional Review Board (IRB-No.E2405/002-021).”

May combine two claims.

VSAs are a human-relevant, vascularized and immune-integrated skin assembloid model for mechanistic studies and preclinical evaluation of pharmacological responses in inflammatory skin diseases.

“Collectively, these findings support VSAs as a human-relevant, vascularized and immune-integrated skin assembloid model for mechanistic studies and preclinical evaluation of pharmacological responses in inflammatory skin diseases.”

Probably not a substantial claim: may not pass S3 (load-bearing).

HDM extract is a major environmental allergen.

“using HDM extract, a major environmental allergen 27,28 .”

• cites [27] Boralevi, F. et al. Epicutaneous aeroallergen sensitization in atopic dermatitis infants — determining the role of epidermal barrier impairment. Allergy 63, 205–210 (2008). sourceNot checked — no full text of the source
• cites [28] Miller, J. D. The role of dust mites in allergy. Clin. Rev. Allergy Immunol. 57, 312–329 (2019). sourceNot checked — no full text of the source

May combine two claims.

Integration of MNCs into VSAs supports immune-cell infiltration and endothelial inflammatory activation upon inflammatory challenge, providing an immune-responsive human skin model for investigating vascular-immune interactions.

“Collectively, these results indicate that integration of MNCs into VSAs supports immune-cell infiltration and endothelial inflammatory activation upon inflammatory challenge, providing an immune-responsive human skin model for investigating vascular-immune interactions.”

May combine two claims.

VSAs display enhanced structural and vascular organization compared with conventional SkOs, providing a more structurally complex vascularized skin assembloid model.

“Collectively, these findings demonstrate that VSAs display enhanced structural and vascular organization compared with conventional SkOs, providing a more structurally complex vascularized skin assembloid model.”

May combine two claims.

HDM stimulation induces STAT6-associated type 2 inflammatory signalling, which is mitigated by upadacitinib and dupilumab.

“Together, these findings suggest that HDM stimulation induces STAT6-associated type 2 inflammatory signalling, which is mitigated by upadacitinib and dupilumab.”

May combine two claims.

Both agents attenuate HDM-induced immune-cell infiltration and alter the spatial distribution of infiltrating immune cells within VSAs.

“This redistribution suggests that both agents attenuate HDM-induced immune-cell infiltration and alter the spatial distribution of infiltrating immune cells within VSAs.”

May combine two claims.

VSAs develop a more mature barrier-associated epidermal phenotype and improved restriction of dye penetration compared with conventional SkOs.

“These results indicate that VSAs develop a more mature barrier-associated epidermal phenotype and improved restriction of dye penetration compared with conventional SkOs.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Upadacitinib is a JAK inhibitor with proven efficacy in moderate-to-severe AD.

“Upadacitinib, a JAK inhibitor with proven efficacy in moderate-tosevere AD 30,31 , and dupilumab, an IL-4Rα antagonist that blocks IL-4/IL-13 signalling 32,33 , were selected.”

• cites [30] Guttman-Yassky, E. et al. Once-daily upadacitinib versus placebo in adolescents and adults with moderate-to-severe atopic dermatitis (Measure Up 1 and Measure Up 2): results from two replicate double-blind, randomised controlled phase 3 trials. Lancet 397, 2151–2168 (2021). sourceNot checked — no full text of the source
• cites [31] Reich, K. et al. Safety and efficacy of upadacitinib in combination with topical corticosteroids in adolescents and adults with moderate-to-severe atopic dermatitis (AD Up): results from a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet 397, 2169–2181 (2021). sourceNot checked — no full text of the source

May combine two claims.

HDM stimulation induces inflammatory remodelling in immune-integrated VSAs with AD-relevant features, characterized by barrier perturbation, epidermal hyperplasia-like remodelling, immune-cell infiltration, myeloid/APC-like activation-associated changes and type 2-associated inflammatory mediator production.

“Together, these findings indicate that HDM stimulation induces inflammatory remodelling in immuneintegrated VSAs with AD-relevant features, characterized by barrier perturbation, epidermal hyperplasia-like remodelling, immune-cell infiltration, myeloid/APC-like activation-associated changes and type 2-associated inflammatory mediator production.”

May combine two claims.

The observed pharmacological responses are consistent with the known mechanisms of clinically used AD therapeutics and support the utility of the VSA platform for preclinical pharmacological evaluation of anti-inflammatory and barrier-modulating agents.

“The observed pharmacological responses are consistent with the known mechanisms of clinically used AD therapeutics and support the utility of the VSA platform for preclinical pharmacological evaluation of anti-inflammatory and barrier-modulating agents.”

May combine two claims.

VSAs capture barrier-associated changes, immune-associated activation, vascular organization and pharmacologically responsive inflammatory readouts within a human-relevant context.

“By capturing barrier-associated changes, immune-associated activation, vascular organization and pharmacologically responsive inflammatory readouts within a human-relevant context, this model provides a useful testbed for studying inflammatory mechanisms”

May combine two claims.

The immune component derived from umbilical cord blood mononuclear cells provides an experimentally tractable immune-integrated system but introduces donor variability and does not fully recapitulate native skin-resident immunity.

“The immune component, derived from umbilical cord blood mononuclear cells, provides an experimentally tractable immuneintegrated system but introduces donor variability and does not fully recapitulate native skin-resident immunity.”

• AssumptionRests on an assumption: The immune component derived from umbilical cord blood mononuclear cells introduces donor variability and does not fully recapitulate native skin-resident immunity.

May combine two claims.

The current VSA model lacks bona fide Langerhans cells, tissue-resident memory T cells, and functionally specialized skin dendritic cell subsets.

“In particular, the current VSA model lacks bona fide Langerhans cells, tissueresident memory T cells, and functionally specialized skin dendritic cell subsets 53 .”

• cites [53] Merad, M., Ginhoux, F. & Collin, M. Origin, homeostasis and function of Langerhans cells and other langerin-expressing dendritic cells. Nat. Rev. Immunol. 8, 935–947 (2008). sourceNot checked — no full text of the source

• AssumptionRests on an assumption: The current VSA platform does not contain bona fide epidermal Langerhans cells.

Probably not a substantial claim: may not pass S3 (load-bearing).

This study was carried out in accordance with the approved guidelines of the Seoul National University Institutional Review Board (IRB-No.2504/003-001 and IRB-No.E2405/002-021).

“This study was carried out in accordance with the approved guidelines of the Seoul National University Institutional Review Board (IRB-No.2504/003-001,and IRB-No.E2405/002-021).”