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Nature CommunicationsCancer BiologyPeer-reviewed journal article

YY1 Enhances the Stability of HIF-1α Protein by Interacting with NUSAP1 in Macrophages within the Prostate Cancer Microenvironment

Wenchao LiiD, Saisai CheniD, Jian Lu, Weipu MaoiD, Shiya ZhengiD, Minhao Zhang, Tiange WuiD, Yurui Chen, Kai Lu, Chunyan Chu, Chuanjun ShuiD, Yue Hou, Xue Yang, Naipeng Shi, Zhijun CheniD, Lihua ZhangiD, Lei ZhangiD, Rong NaiD, Ming CheniD, Shenghong JuiD, Dingxiao ZhangiD, Yi MaiD, Bin XuiD

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Abstract

Immune checkpoint therapy for prostate cancer (PCa) has failed in clinical trials; however, the precise underlying mechanisms involved remain elusive. PCa, a classic "immune-cold" tumor, is characterized by an immunosuppressive tumor microenvironment. Within this milieu, macrophages, the predominant immune cell population, have a propensity to infiltrate the hypoxic zones of tumors. In a previous study, we showed that Yin Yang 1 (YY1) is highly expressed in macrophages in PCa tissues. Here, through multiplexed imaging mass cytometry (IMC) of a PCa tissue microarray, we further demonstrate that YY1+ macrophages aggregate in hypoxic areas of tumors and that hypoxia promotes the phase separation of YY1 in the nucleus by increasing YY1 tyrosine phosphorylation in macrophages. Furthermore, YY1 binds to NUSAP1 and promotes the SUMOylation of HIF-1, which promotes phase separation and stabilization of the HIF-1 protein. We also demonstrated that either treatment with a small molecule inhibitor (tenapanor) to decrease the YY1-NUSAP1-HIF-1 interaction or myeloid-specific YY1 gene knockout impairs subcutaneous PCa tumor formation. Furthermore, we present a first-generation tetrahedral DNA nanostructure (TDN) based on the proteolysis targeting chimera (PROTAC) technique, named YY1-DcTAC, which targets and degrades YY1 in tumor-associated macrophages. In a PCa mouse model, YY1-DcTAC exhibited prolonged drug efficacy, robust macrophage-specific responsiveness, potent antitumor effects, and increased CD8+ T cell tumor infiltration. In summary, our findings underscore the pivotal role of YY1 within the hypoxia/HIF-1 pathway in tumor-associated macrophages and affirm the therapeutic potential of targeting YY1 for treating PCa.

In-depth summary

Claude Sonnet 4.6 · xhigh effort · 8 Nov 2026 · from the full text of version 1 (8 Nov 2024) · CC BY 4.0

In THP-1 cells, hypoxia drives Lyn-dependent YY1 phosphorylation and phase separation; YY1 and NUSAP1 SUMOylate and stabilize HIF-1α, and degrading YY1 curbs mouse prostate tumors.

Prostate cancer is an immune-cold tumor that responds poorly to checkpoint therapy, and YY1 has structural heterogeneity and few active sites, which hinders drug development. This work links macrophage YY1 to hypoxia and HIF-1α stability, and tests tenapanor and a PROTAC-based DNA nanostructure as ways to target it.

Key results

  1. ShownYY1-positive macrophages were enriched in HIF-1α-positive hypoxic patches of prostate tumors, but not in hypoxic patches of adjacent noncancerous tissue or in HIF-1α-negative areas.

    Imaging mass cytometry of 46 PCa and 26 adjacent noncancerous tissues; a greater percentage of YY1+ than YY1- macrophages in tumor hypoxic regions (P < 0.05); no substantial difference in adjacent tissue or HIF-1α-negative patches. · YY1+ macrophages accumulate in hypoxic PCa…; Figure legends

  2. ShownIn THP-1 cells, CoCl2 hypoxia triggers nuclear YY1 phase separation through Lyn-dependent tyrosine phosphorylation within the YY1 disordered region, without changing YY1 abundance.

    YY1 level unchanged across CoCl2 doses and durations; p-Tyr but not p-Ser rose; the Y145F/Y185F/Y251F/Y254F mutant blocked the p-Tyr rise and the Y8F/Y383F mutant did not; SU6656 and Lyn siRNA reduced droplets; reversal after 120 min reoxygenation. · Hypoxia enhances YY1 phase separation by…; Figure legends

  3. SuggestedYY1 binds NUSAP1, whose SAP domain mediates SUMOylation of HIF-1α, which reduces HIF-1α degradation and favors HIF-1α condensates; K391 appears more important than K477.

    YY1 knockdown lowered HIF-1α protein with unchanged mRNA and shortened its stability in a CHX chase; NUSAP1 knockdown lowered HIF-1α and its SUMOylation; the ΔSAP variant failed to raise SUMOylation; binding mapped to NUSAP1 240–250 and YY1 200–220; K391F reduced SUMOylation. · YY1 stabilizes HIF-1α by inhibiting its…; NUSAP1 promotes HIF-1α SUMOylation and inhibits…; HIF-1α undergoes SUMOylation-related phase…

  4. SuggestedTenapanor, picked by docking as a disruptor of the YY1–NUSAP1–HIF-1α complex, slowed subcutaneous RM-1 tumor growth and increased CD8+ T cell infiltration when delivered with a macrophage-targeting peptide liposome.

    Docking of 1729 small molecules; tenapanor in the top 10 docking scores for all three proteins; TEN-M2pep slowed tumor growth vs control and raised CD8+ T cell density (IHC, flow); with PD-1 inhibitor, tumors grew significantly more slowly; exact P values and n not given. · Targeting the YY1–NUSAP1–HIF-1α nexus in…; Figure legends

  5. SuggestedDegrading YY1 in tumor macrophages, with a CD206-responsive tetrahedral DNA PROTAC (YY1-DcTACs) or myeloid-specific YY1 knockout, restrained subcutaneous tumor growth and raised CD8+ T cell proportions.

    CD3+CD8+ T cells 5.38% ± 4.78% vs 1.46% ± 0.11% in YY1-DcTACs vs control tumors; tumor inhibition until day 14; knockout mice showed slower growth at 21 days and a higher CD8+ to CD3+ ratio; sample sizes not stated. · Construction and application of YY1-DbTACs and…; Figure legends

Methods, data and reproducibility

Imaging mass cytometry of a prostate tissue microarray, then THP-1 cell assays (CoCl2 hypoxia, co-IP, ubiquitination and SUMOylation assays, phase separation imaging), docking-based drug screening, and subcutaneous mouse tumor models testing tenapanor, DNA-based PROTACs and myeloid YY1 knockout.

  • Prostate tissue microarray of radical prostatectomy specimens, tumor cores and adjacent noncancerous cores, analyzed by imaging mass cytometry · human · n = 46 PCa and 26 adjacent noncancerous tissues
  • THP-1 monocyte/macrophage cell experiments under CoCl2 hypoxia (co-IP, imaging, ubiquitination and SUMOylation assays) · human
  • RNA-seq of THP-1 cells under hypoxia or YY1 overexpression, and IP-mass spectrometry of YY1 in THP-1 cells · human
  • Docking screen of small molecules against YY1, NUSAP1 and HIF-1α · n = 1729 small molecules
  • Subcutaneous RM-1 prostate tumor models in C57BL/6 male mice (tenapanor liposomes, DNA PROTACs, PD-1 inhibitor, OT-1 T cell combinations) · mouse
  • Myeloid-specific YY1 conditional knockout mice (Lyz2-Cre, flox/flox) vs wild-type, subcutaneous tumor model · mouse
  • RNA-seq of harvested tenapanor-treated subcutaneous tumors · mouse
Methods & tools
Hyperion Imaging System (Fluidigm), PONDR (VSL2), SUMOplot, Discovery Studio, AlphaFold, HDOCK, Rosetta, GSEA, FlowJo V10, SPSS 16.0, Patch and milieu detection algorithm, FRAP assay
Reagents & environment
SPSS software version 16.0; FlowJo V10; no containers or code environment described.
Reproducibility gaps
Data statement says all data are in the main text or supplement; no repository accession or code link is given. Table S2 (siRNA sequences) is a supplementary Excel file.
Limitations (4)
  • Each YY1-DcTACs molecule releases only one DbTAC molecule, so drug loading capacity is limited and constrains therapeutic potential. · stated by the authors
  • The double-stranded DNA of YY1-DbTACs is degraded by nucleases within 24 h, requiring frequent dosing, which motivated the tetrahedral design. · stated by the authors
  • Hypoxia was modeled mainly with CoCl2 (a chemical mimetic) in THP-1 cells and efficacy in subcutaneous RM-1 tumors; tenapanor's mechanism rests on docking and IP, and off-target effects of this NHE3 inhibitor were not excluded. · noted in this summary
  • Mouse group sizes, effect sizes and exact P values are mostly not reported, and the text and the Figure 4D legend disagree on the direction of HIF-1α ubiquitination after NUSAP1 knockdown. · noted in this summary

Read this if you study hypoxia, HIF-1α regulation or macrophage-targeted therapy in immune-cold tumors, or PROTAC-style DNA nanostructures for transcription factors.

Citations
28
FWCI
9.51
Field percentile
top 1%

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