Acute pharmacological degradation of Helios destabilizes regulatory T cells
Open accessAbstract
The zinc-finger transcription factor Helios is critical for maintaining the identity, anergic phenotype and suppressive activity of regulatory T (T reg) cells. While it is an attractive target to enhance the efficacy of currently approved immunotherapies, no existing approaches can directly modulate Helios activity or abundance. Here, we report the structure-guided development of small molecules that recruit the E3 ubiquitin ligase substrate receptor cereblon to Helios, thereby promoting its degradation. Pharmacological Helios degradation destabilized the anergic phenotype and reduced the suppressive activity of T reg cells, establishing a route towards Helios-targeting therapeutics. More generally, this study provides a framework for the development of small-molecule degraders for previously unligandable targets by reprogramming E3 ligase substrate specificity.
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Written from the paper’s full text.
TL;DR
The authors developed CRBN-recruiting small-molecule degraders ALV1 and ALV2, with ALV2 preferentially degrading Helios (IKZF2) and destabilizing regulatory T cells. Acute Helios degradation recapitulated genetic Ikzf2 loss, reducing Treg anergy and suppressive activity in murine and human systems.
Key findings
- ALV1 and ALV2 bound CRBN and induced CRBN-Helios dimerization; ALV1 degraded both Ikaros and Helios, while ALV2 preferentially degraded Helios, and neither degraded GSPT1. In Jurkat proteomics across ~7,900 proteins, ALV1 showed similar IKZF1/2/3 degradation, ALV2 preferentially promoted Helios degradation, and ZNF324 was identified as a potential novel ALV2 target.
- The crystal structure of DDB1ΔB-CRBN-ALV1-IKZF2ZnF2 was determined at 3.8 Å; H141 of IKZF2 was ~3.5 Å from the ALV1 ethylbenzene moiety, and ALV1’s tail would clash with GSPT1 V536/Q534, providing a structural rationale for lack of GSPT1 degradation.
- In CrbnI391V/I391V mice, ALV2 (100 mg/kg i.p. twice daily for 7 days) reduced Helios but not Ikaros in splenic CD4+ FoxP3+ Tregs; ex vivo ALV2-treated murine FoxP3+ Tregs had lower Helios and a significantly higher percentage of IFNγ-producing Tregs after PMA/ionomycin.
- In human Tregs, ALV2 degraded Helios/Eos; proteomics across ~6,800 proteins showed preferential IKZF2/4 degradation and ZMYM2 reduction, and ALV2 increased IL-2- or IFNγ-producing Tregs and reduced suppression of responder cell proliferation. ALV1 and ALV2 also promoted IL-2 secretion in Jurkat cells to a greater extent than lenalidomide.
Methods
- Structure-guided medicinal chemistry with TR-FRET CRBN binding and CRBN-Helios dimerization assays, EGFP-P2A-mCherry reporter degradation assays, immunoblotting, X-ray crystallography, and multiplexed TMT LC-MS3 proteomics.
- Cell systems included Jurkat cells, human Tregs expanded ex vivo, and murine Tregs from wildtype or CrbnI391V/I391V mice; readouts included flow cytometry, IL-2 ELISA, and a human Treg suppression assay.
- In vivo pharmacodynamics used CrbnI391V/I391V mice treated with vehicle or ALV2 (100 mg/kg i.p. twice daily for 7 days), with spleens collected 4 h after the final treatment.
Limitations
- Stated: the 3.8 Å structure resolution prevents detailed analysis; detailed drivers of selectivity require further structures with IKZF1/3; the ZNF324 degron remains unidentified; and the absence of mouse models that recapitulate human imide immunomodulatory pharmacology led the authors to rely on ex vivo human Tregs.
- Stated/evident: ALV2 also degrades Eos (IKZF4) and ZMYM2, so it is not Helios-exclusive, and proteomics identified additional degradation targets.
- Evident from the text: the in vivo study reports pharmacodynamic Helios reduction in splenic Tregs, but no tumor efficacy, autoimmunity, or broader safety data are shown.
Model-written summary. Check it against the paper before relying on it.
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Claims the paper makes
0 of 6 cited sources checked · 0 supported · 0 flagged
- Findingown data
The structure of the DDB1-CRBN-IKZF2ZnF2-ALV1 complex was determined to 3.8 Å resolution.
“Crystals were obtained for the 152.8 kDa complex, and its structure was determined to 3.8 Å resolution (Fig. 2a, Supplementary Table 2).”
Quote in the paperSubstantial claim - Findingown data
Novel imide analogs with anilinomaleimide cores bind CRBN in cells.
“Novel imide analogs with anilinomaleimide cores bind CRBN in cells to induce Helios degradation”
Quote in the paperSubstantial claim - Findingown data
PMA/ionomycin restimulation of human Tregs expanded in ALV2 significantly increased the percentage of IL-2 or IFNγ-producing Tregs over vehicle control.
“Consistent with the loss of Treg anergy observed in murine cells, PMA/ionomycin restimulation of human Tregs expanded in the presence of ALV2 significantly increased the percentage of IL-2 or IFNγ-producing Tregs over vehicle control (Fig. 4a, b).”
Quote in the paperSubstantial claim - Findingown data
The glycine residue indicated by the red arrow is present at least once in all downregulated proteins.
“and is present at least once in all downregulated proteins.”
Quote in the paperNot a substantial claim - Findingown data
Novel imide analogs with anilinomaleimide cores induce Helios degradation.
“Novel imide analogs with anilinomaleimide cores bind CRBN in cells to induce Helios degradation”
Quote in the paperSubstantial claim
- Findingown data
Small molecules were developed that recruit CRBN to Helios, promoting its degradation.
“Here, we report the structure-guided development of small molecules that recruit the E3 ubiquitin ligase substrate receptor Cereblon to Helios, thereby promoting its degradation.”
Quote in the paperSubstantial claim- Rests on an assumption: No existing approaches can directly modulate Helios activity or abundance, and no Helios-targeting small molecules have been reported.
- Findingown data
Both ALV1 and ALV2 promoted IL-2 secretion to a greater extent than lenalidomide.
“Consistent with the ascribed role of Helios in suppressing IL-2, both ALV1 and ALV2 promoted IL-2 secretion to a greater extent than lenalidomide, suggesting that Helios more strongly represses IL-2 production than Ikaros/Aiolos in this context.”
Quote in the paperSubstantial claim - Findingown data
Human Tregs expanded in the presence of ALV2 were less able to repress the proliferation of responder cells.
“We also found that human Tregs expanded in the presence of ALV2 were less able to repress the proliferation of responder cells (Fig. 4c).”
Quote in the paperSubstantial claim - Interpretationcitation
Acute pharmacological degradation of Helios recapitulates the phenotype of genetic loss of Ikzf2 in Tregs.
“Notably, the preferential Helios degrader ALV2 destabilized the anergic phenotype of regulatory Tregs, consistent with previously reported phenotypes of Ikzf2−/− Tregs5,6,29, demonstrating that acute pharmacological degradation of Helios recapitulates the phenotype of genetic loss of Ikzf2 in Tregs.”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [5] Nakagawa, H. et al. Instability of Helios-deficient Tregs is associated with conversion to a T-effector phenotype and enhanced antitumor immunity. Proc. Natl Acad. Sci. USA 113, 6248–6253 (2016). source
Not checked in the free run.
- cites [6] Kim, H. J. et al. Stable inhibitory activity of regulatory T cells requires the transcription factor Helios. Science 350, 334–339 (2015). source
Not checked in the free run.
- cites [29] Sebastian, M. et al. Helios controls a limited subset of regulatory T cell functions. J. Immunol. 196, 144–155 (2016). source
Not checked in the free run.
- Interpretationnone given
The glycine residue indicated by the red arrow is a key determinant of imide-dependent degradation.
“The glycine residue indicated by the red arrow is a key determinant of imide dependent degradation”
Quote in the paperNot a substantial claim - Interpretationown data
The critical histidine residue in Helios regulates sensitivity to imide-induced degradation.
“Critical histidine residue in Helios regulates sensitivity to imide-induced degradation”
Quote in the paperSubstantial claim - Interpretationown data
Acute Helios degradation destabilizes murine Tregs.
“Acute Helios degradation in destabilizes murine Tregs”
Quote in the paperSubstantial claim - Interpretationown data
Helios degradation promotes IL-2 secretion.
“Helios degradation promotes IL-2 secretion”
Quote in the paperNot a substantial claim
- May combine two claims. 14
- The quote only partly states this claim. 1
- Classified as probably not a substantial claim. 4
May combine two claims.
Findingown dataALV2 treatment reduced Helios but not Ikaros levels in splenic CD4+ FoxP3+ Tregs in vivo.
“Finally, we administered ALV2 or vehicle control to CrbnI391V/I391V mice and found that ALV2 treatment reduced Helios but not Ikaros levels in splenic CD4+ FoxP3+ Tregs (Extended Data Fig. 6c, Supplementary Table 3), demonstrating that ALV2 can induce selective Helios degradation in vivo.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataNeither ALV1 nor ALV2 destabilized GSPT1.
“Neither ALV1 nor ALV2 destabilized GSPT1, and as expected, lenalidomide only induced degradation of Ikaros, while CC-885 induced degradation of both Ikaros and GSPT1 (Extended Data Fig 2c).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataALV1 and ALV2 bound CRBN in vitro with greater potency than lenalidomide but less than CC-885, and induced CRBN-Helios dimerization in the TR-FRET assay.
“Both ALV1 and ALV2 bound CRBN in vitro with greater potency than lenalidomide (4) but less than CC-885 (5) (Fig. 1b), and both compounds also induced CRBN-Helios dimerization in the TR-FRET assay, with greater activity than either CC-885 (positive control) or lenalidomide (negative control) (Fig. 1c).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataALV1 degraded both Ikaros and Helios, while ALV2 exhibited relative selectivity for Helios.
“Our medicinal chemistry campaign optimized for compounds with distinct pharmacological degradation profiles, resulted in the development of ALV1, which degraded both Ikaros and Helios, and ALV2, which exhibited relative selectivity for Helios.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataThe residue that controls sensitivity to IMiD-induced degradation is glutamine in IKZF1/3 and histidine in IKZF2/4.
“with the residue that controls sensitivity to IMiD-induced degradation (glutamine residue in IKZF1/3, histidine in IKZF2/4) highlighted in cyan”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataProteome-wide analysis revealed that ALV1 exhibited similar degradation activity towards IKZF1/2/3 while ALV2 preferentially promoted Helios degradation.
“Thus, proteome-wide analysis revealed that the nonselective compound ALV1 exhibited similar degradation activity towards IKZF1/2/3 while ALV2 preferentially promoted Helios degradation, as also observed by immunoblot.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataPharmacological Helios degradation destabilized the anergic phenotype and reduced the suppressive activity of regulatory T cells.
“Pharmacological Helios degradation destabilized the anergic phenotype and reduced the suppressive activity of regulatory T cells, establishing a route towards Helios-targeting therapeutics.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataIn human Tregs, ALV1 potently induced degradation of both Ikaros and Helios, while ALV2 selectively destabilized Helios.
“Consistent with our previous results, ALV1 potently induced degradation of both Ikaros and Helios, while ALV2 selectively destabilized Helios (Fig. 3a).”
Quote in the paperSubstantial claimMay combine two claims. The quote only partly states this claim.
Findingown dataALV2 treatment reduced Helios levels and increased the percentage of IFNγ-producing Tregs in CrbnI391V/I391V murine Tregs.
“Moreover, restimulation of cells with PMA/ionomycin induced a significantly higher percentage of IFNγ-producing Tregs in ALV2-treated compared to vehicle-treated Foxp3+ Tregs (Extended Data Fig. 6b), consistent with Ikzf2−/− Tregs acquiring effector functions, which demonstrates that acute pharmacologic degradation of Helios can recapitulate its genetic loss in murine Tregs.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Findingown dataThe DDB1ΔB-CRBN-ALV1-IKZF2ZnF2 ternary complex crystallized in space group P21 with four complexes in the asymmetric unit.
“The DDB1ΔB-CRBN-ALV1- IKZF2ZnF2 ternary complex crystallized in space group P21 with four complexes in the asymmetric unit.”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataAcute Helios degradation occurs in CrbnI391V/I391V but not wildtype murine Tregs.
“Acute Helios degradation in CrbnI391V/I391V but not wildtype murine Tregs”
Quote in the paperSubstantial claimClassified as probably not a substantial claim. May combine two claims.
InterpretationreasoningThe study provides a roadmap for the development of Helios-degrading therapeutics as immunotherapy agents that may synergize with immune checkpoint blockade.
“Thus, our data suggest that Helios-mediated regulation of Treg stability is not established during thymic and/or peripheral Treg development, and provides a roadmap for the development of Helios-degrading therapeutics as immunotherapy agents that may synergize with immune checkpoint blockade.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing), S4 (based).
Interpretationnone givenLoss of Helios abundance may be secondary to defects in translation and subsequent initiation of programmed cell death.
“Loss of Helios abundance may be secondary to defects in translation and subsequent initiation of programmed cell death.”
Quote in the paperNot a substantial claimMay combine two claims.
PremisecitationHelios is critical for maintaining the identity, anergic phenotype, and suppressive activity of regulatory T cells.
“The zinc finger transcription factor Helios is critical for maintaining the identity, anergic phenotype, and suppressive activity of regulatory T cells.”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [5] Nakagawa, H. et al. Instability of Helios-deficient Tregs is associated with conversion to a T-effector phenotype and enhanced antitumor immunity. Proc. Natl Acad. Sci. USA 113, 6248–6253 (2016). source
Not checked in the free run.
- cites [6] Kim, H. J. et al. Stable inhibitory activity of regulatory T cells requires the transcription factor Helios. Science 350, 334–339 (2015). source
Not checked in the free run.
- cites [7] Yates, K., Bi, K., Haining, W. N., Cantor, H. & Kim, H. J. Comparative transcriptome analysis reveals distinct genetic modules associated with Helios expression in intratumoral regulatory T cells. Proc. Natl. Acad. Sci. USA 115, 2162–2167 (2018). source
Not checked in the free run.
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Study design and conductown dataProtein geometry analysis revealed 0.60% Ramachandran outliers, with 95.72% residues in favored regions and 3.68% residues in allowed regions for the complex with ALV1.
“Protein geometry analysis revealed 0.60%, Ramachandran outliers, with 95.72% residues in favored regions and 3.68% residues in allowed regions for the complex with ALV1.”
Quote in the paperNot a substantial claim
The paper relies on this without support.
No existing approaches can directly modulate Helios activity or abundance, and no Helios-targeting small molecules have been reported.
“While it is an attractive target to enhance the efficacy of currently approved immunotherapies, no existing approaches can directly modulate Helios activity or abundance.”
“While Helios is an attractive immune-oncology target, transcription factors are traditionally considered to be hard to drug, and no Helios-targeting small molecules have been reported.”
The paper relies on this without support.
Phenotypic destabilization of Tregs caused by Helios deficiency preferentially occurs in tumor-infiltrating Tregs.
“This phenotypic destabilization preferentially occurs in tumor-infiltrating Tregs, which suggests that pharmacologically targeting Helios may mitigate the risk of autoimmunity.”
The paper relies on this without support.
Transcription factors are traditionally considered hard to drug.
“While Helios is an attractive immune-oncology target, transcription factors are traditionally considered to be hard to drug, and no Helios-targeting small molecules have been reported.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
MGs bind to the C-terminal domain of cereblon and induce conformational changes that create a new surface interacting with zinc fingers containing a beta-turn CXXCG motif.
“With respect to the molecular glue-induced interaction of zinc finger transcription factors with cereblon, structural studies show that MGs bind to the C-terminal domain of cereblon and induce conformational changes that result in a new surface that interacts with zinc fingers containing a beta turn motif with sequence CXXCG7–9.”
From Structural and biophysical comparisons of the pomalidomide- and CC-220-induced interactions of SALL4 with cereblon (2023) · cites this paper as [9]
The Results section, “Structural basis of ALV1 induced Helios/CRBN dimerization” (page unavailable), shows ALV1 bound to CRBN and a zinc finger interacting with its C-terminal domain through a β-hairpin loop. The Discussion (page unavailable) identifies CxxCG as a motif permissive of degradation. The paper observes CRBN in a closed conformation but does not establish that molecular-glue binding induces a conformational change that creates a new interaction surface. (LLM, conf=0.86)
Experimental — accuracy not yet measuredIt is the third binding mode for ZFs to CRBN:MG complexes.
“Additionally, it is the third binding mode for ZFs to CRBN:MG complexes adding to our understanding of the binding of single zinc fingers such as IKZF13,7 and ZNF6923, and the binding of two zinc fingers including C-terminal ancillary ZF3 in the IKZF2 ZF2-3 complex9,10.”
From Structural and biophysical comparisons of the pomalidomide- and CC-220-induced interactions of SALL4 with cereblon (2023) · cites this paper as [9]
The Results section (page unavailable) describes a binding mode for a single IKZF2 zinc finger, ZnF2, in a CRBN–ALV1 complex. It does not establish that this is the third binding mode or describe binding by the two-finger IKZF2 ZF2–3 complex with an ancillary ZF3. (LLM, conf=0.86)
Experimental — accuracy not yet measuredRational structure-guided design led to flexible imide analogues that selectively degrade IKZF2 and the structurally related IKZF4.
“Rational structure-guided design led to the development of flexible imide analogues that selectively degrade iKZF2 and the structurally related iKZF4.”
From From serendipity to strategy: molecular glue degraders in cancer therapeutics (2025) · cites this paper as wang et al. 2021
TR-FRET and fluorescence polarization assays offer robust scalable formats for detecting molecular glue-induced interactions in vitro.
“Similarly, tR-FRet and fluorescence polarization (FP) assays offer robust scalable formats for detecting molecular glue-induced interactions in vitro (Simonetta et al. 2019;wang et al. 2021;Hsia et al. 2024;Kozicka et al. 2024).”
From From serendipity to strategy: molecular glue degraders in cancer therapeutics (2025) · cites this paper as wang et al. 2021
The Experimental Materials and Methods sections on cellular degradation and CRBN engagement describe GFP/mCherry imaging assays in cells, not in-vitro assays for molecular glue-induced interactions. Searches for TR-FRET, time-resolved fluorescence energy transfer, and fluorescence polarization found no passage in our copy establishing that those assays provide robust, scalable formats. Page numbers were not available. (LLM, conf=0.75)
Experimental — accuracy not yet measuredSeveral rounds of optimization led to ALV1 and ALV2.
“Several rounds of optimization led to ALV1 and ALV2 (Fig. 7 and 8).”
From Chasing molecular glue degraders: screening approaches (2022) · cites this paper as [167]
In 2021, Wang et al. reported structure-guided development of CRBN-dependent MG degraders of Helios (IKZF2), a transcription factor involved in immunosuppression.
“In 2021, Wang et al. reported the structure-guided development of CRBN-dependent MG degraders of Helios (IKZF2), a transcription factor involved in immunosuppression.167”
From Chasing molecular glue degraders: screening approaches (2022) · cites this paper as [167]
ALV1 induces CRBN-dependent degradation of Helios and retains Ikaros destabilization, while ALV2 has relative selectivity for Helios.
“The former induces CRBN-dependent degradation of Helios and also retains Ikaros destabilization, while the latter has relative selectivity for Helios.”
From Chasing molecular glue degraders: screening approaches (2022) · cites this paper as [167]
Wang et al. synthesized the IMiD derivative ALV-02-146-03 and checked that it could engage CRBN.
“First, Wang et al. synthesized the IMiD derivative ALV-02-146-03 and checked that it could engage CRBN.”
From Chasing molecular glue degraders: screening approaches (2022) · cites this paper as [167]
Eos (IKZF4) also encodes a histidine residue at the same position as Helios, hence ALV1 and ALV2 also induce Eos destabilization.
“Of note, Eos (IKZF4) also encodes a histidine residue at the same position as Helios, hence ALV1 and ALV2 also induce Eos destabilization.167”
From Chasing molecular glue degraders: screening approaches (2022) · cites this paper as [167]
Helios degradation weakened the anergic phenotype and reduced suppressive activity of Treg cells.
“Helios degradation weakened the anergic phenotype (T cell unresponsiveness) and reduced the suppressive activity of Treg cells.”
From Chasing molecular glue degraders: screening approaches (2022) · cites this paper as [167]
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“Indeed, thalidomide analogs (also known as IMiDs, or immunomodulatory imide drugs) can induce the degradation of a variety of proteins, such as Ikaros (IKZF1) and Aiolos (IKZF3) 8,9 , casein kinase 1 α (CK1 α ) 10 , ZFP91 11 , SALL4 12,13 , and GSPT1 14 .”
Self-citationStable inhibitory activity of regulatory T cells requires the transcription factor Helios
Kim, H.-J., Barnitz, R. A., Kreslavsky, T., Brown, F. D., Moffett, H., Lemieux, M. E., Kaygusuz, Y., Meissner, T., Holderried, T. A. W., Chan, S., et al. (2015) · Science · doi:10.1126/science.aad0616 · Cited by 441 · see citing works
Mapping the Degradable Kinome Provides a Resource for Expedited Degrader Development
Donovan, K. A., Ferguson, F. M., Bushman, J. W., Eleuteri, N. A., Bhunia, D., Ryu, S., Tan, L., Shi, K., Yue, H., Liu, X., et al. (2020) · Cell · doi:10.1016/j.cell.2020.10.038 · Cited by 359 · see citing works
Self-citationSALL4 mediates teratogenicity as a thalidomide-dependent cereblon substrate
Matyskiela, M. E., Couto, S., Zheng, X., Lu, G., Hui, J., Stamp, K., Drew, C., Ren, Y., Wang, M., Carpenter, A., et al. (2018) · Nature Chemical Biology · doi:10.1038/s41589-018-0129-x · Cited by 338 · see citing works
Eos Mediates Foxp3-Dependent Gene Silencing in CD4 + Regulatory T Cells
Pan, F., Yu, H., Dang, E. V., Barbi, J. J., Pan, X., Grosso, J. F., Jinasena, D., Sharma, S. M., McCadden, E., Getnet, D., et al. (2009) · Science · doi:10.1126/science.1176077 · Cited by 335 · see citing works
The Ikaros gene family: Transcriptional regulators of hematopoiesis and immunity
John, L. B., & Ward, A. C. (2011) · Molecular Immunology · doi:10.1016/j.molimm.2011.03.006 · Cited by 231 · see citing works
Cullin–RING ubiquitin E3 ligase regulation by the COP9 signalosome
Cavadini, S., Fischer, E. S., Bunker, R. D., Potenza, A., Lingaraju, G. M., Goldie, K. N., Mohamed, W. I., Faty, M., Petzold, G., Beckwith, R. E. J., et al. (2016) · Nature · doi:10.1038/nature17416 · Cited by 213 · see citing works
Self-citationNakagawa, H., Sido, J. M., Reyes, E. E., Kiers, V., Cantor, H., & Kim, H.-J. (2016) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1604765113 · Cited by 176 · see citing works
pSILAC mass spectrometry reveals ZFP91 as IMiD-dependent substrate of the CRL4CRBN ubiquitin ligase
An, J., Ponthier, C. M., Sack, R., Seebacher, J., Stadler, M. B., Donovan, K. A., & Fischer, E. S. (2017) · Nature Communications · doi:10.1038/ncomms15398 · Cited by 176 · see citing works
Self-citationHelios Controls a Limited Subset of Regulatory T Cell Functions
Sebastian, M., Lopez-Ocasio, M., Metidji, A., Rieder, S. A., Shevach, E. M., & Thornton, A. M. (2016) · The Journal of Immunology · doi:10.4049/jimmunol.1501704 · Cited by 172 · see citing works
Crbn I391V is sufficient to confer in vivo sensitivity to thalidomide and its derivatives in mice
Fink, E. C., McConkey, M., Adams, D. N., Haldar, S. D., Kennedy, J. A., Guirguis, A. A., Udeshi, N. D., Mani, D. R., Chen, M., Liddicoat, B., et al. (2018) · Blood · doi:10.1182/blood-2018-05-852798 · Cited by 131 · see citing works
Helios Induces Epigenetic Silencing of <i>Il2</i> Gene Expression in Regulatory T Cells
Baine, I., Basu, S., Ames, R., Sellers, R. S., & Macian, F. (2013) · The Journal of Immunology · doi:10.4049/jimmunol.1200792 · Cited by 116 · see citing works
Selective Degradation of GSPT1 by Cereblon Modulators Identified via a Focused Combinatorial Library
Powell, C. E., Du, G., Che, J., He, Z., Donovan, K. A., Yue, H., Wang, E. S., Nowak, R. P., Zhang, T., Fischer, E. S., et al. (2020) · ACS Chemical Biology · doi:10.1021/acschembio.0c00520 · Cited by 88 · see citing works
Self-citationAbdulrahman, W., Uhring, M., Kolb-Cheynel, I., Garnier, J., Moras, D., Rochel, N., Busso, D., & Poterszman, A. (2009) · Analytical Biochemistry · doi:10.1016/j.ab.2008.10.044 · Cited by 82 · see citing works
Gokhale, A. S., Gangaplara, A., Lopez-Ocasio, M. D., Thornton, A. M., & Shevach, E. M. (2019) · Journal of Autoimmunity · doi:10.1016/j.jaut.2019.06.011 · Cited by 66 · see citing works
The Ikaros family of zinc-finger proteins
Fan, Y., & Lu, D. (2016) · Acta Pharmaceutica Sinica B · doi:10.1016/j.apsb.2016.06.002 · Cited by 61 · see citing works
Yates, K., Bi, K., Haining, W. N., Cantor, H., & Kim, H.-J. (2018) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1720447115 · Cited by 55 · see citing works
Phasertng: directed acyclic graphs for crystallographic phasing
McCoy, A. J., Stockwell, D. H., Sammito, M. D., Oeffner, R. D., Hatti, K. S., Croll, T. I., & Read, R. J. (2020) · Acta Crystallographica Section D Structural Biology · doi:10.1107/s2059798320014746 · Cited by 13 · see citing works
Phasertng: directed acyclic graphs for crystallographic phasing.
McCoy, A. J., Stockwell, D. H., Sammito, M. D., et al. (2021) · Apollo (University of Cambridge) · doi:10.17863/cam.63804 · Cited by 1 · see citing works
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Molecular glue CELMoD compounds are regulators of cereblon conformation
Watson, E. R., Novick, S., Matyskiela, M. E., Chamberlain, P. P., H. de la Peña, A., Zhu, J., Tran, E., Griffin, P. R., Wertz, I. E., & Lander, G. C. (2022) · Science · doi:10.1126/science.add7574 · Cited by 208 · see citing works
Chasing molecular glue degraders: screening approaches
Domostegui, A., Nieto-Barrado, L., Perez-Lopez, C., & Mayor-Ruiz, C. (2022) · Chemical Society Reviews · doi:10.1039/d2cs00197g · Cited by 200 · see citing works
From Thalidomide to Rational Molecular Glue Design for Targeted Protein Degradation
Oleinikovas, V., Gainza, P., Ryckmans, T., Fasching, B., & Thomä, N. H. (2024) · Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev-pharmtox-022123-104147 · Cited by 178 · see citing works
Discovery and characterization of a selective IKZF2 glue degrader for cancer immunotherapy
Bonazzi, S., d’Hennezel, E., Beckwith, R. E. J., Xu, L., Fazal, A., Magracheva, A., Ramesh, R., Cernijenko, A., Antonakos, B., Bhang, H.-E. C., et al. (2023) · Cell Chemical Biology · doi:10.1016/j.chembiol.2023.02.005 · Cited by 167 · see citing works
Mining the CRBN target space redefines rules for molecular glue-induced neosubstrate recognition
Petzold, G., Gainza, P., Annunziato, S., Lamberto, I., Trenh, P., McAllister, L. A., DeMarco, B., Schwander, L., Bunker, R. D., Zlotosch, M., et al. (2025) · Science · doi:10.1126/science.adt6736 · Cited by 117 · see citing works
Razumkov, H., Jiang, Z., Baek, K., You, I., Geng, Q., Donovan, K. A., Tang, M. T., Metivier, R. J., Mageed, N., Seo, P., et al. (2024) · Journal of the American Chemical Society · doi:10.1021/jacs.4c06127 · Cited by 53 · see citing works
Self-citationDesign of a Cereblon construct for crystallographic and biophysical studies of protein degraders
Kroupova, A., Spiteri, V. A., Rutter, Z. J., Furihata, H., Darren, D., Ramachandran, S., Chakraborti, S., Haubrich, K., Pethe, J., Gonzales, D., et al. (2024) · Nature Communications · doi:10.1038/s41467-024-52871-9 · Cited by 47 · see citing works
Ma, X., Leon, B., Ornelas, E., Dovala, D., Tandeske, L., Luu, C., Pardee, G., Widger, S., Solomon, J. M., Beckwith, R. E. J., et al. (2023) · Scientific Reports · doi:10.1038/s41598-023-48606-3 · Cited by 21 · see citing works
“Additionally, it is the third binding mode for ZFs to CRBN:MG complexes adding to our understanding of the binding of single zinc fingers such as IKZF1 3,7 and ZNF692 3 , and the binding of two zinc fingers including C-terminal ancillary ZF3 in the IKZF2 ZF2-3 complex 9,10 .”
BackgroundFrom serendipity to strategy: molecular glue degraders in cancer therapeutics
Darling, S., Bajrami, I., & West, S. C. (2025) · Critical Reviews in Biochemistry and Molecular Biology · doi:10.1080/10409238.2025.2564068 · Cited by 10 · see citing works
“Similarly, tR-FRet and fluorescence polarization (FP) assays offer robust scalable formats for detecting molecular glue-induced interactions in vitro (Simonetta et al. 2019; wang et al. 2021; Hsia et al. 2024; Kozicka et al. 2024).”
- Authors
- Wang, E. S., Verano, A. L., Nowak, R. P., Yuan, J. C., Donovan, K. A., Eleuteri, N. A., Yue, H., Ngo, K. H., Lizotte, P. H., Gokhale, P. C., Gray, N. S., & Fischer, E. S.
- Affiliations
- Harvard University (US)
- Dana-Farber Cancer Institute (US)
- Corresponding author
- Gray, Nathanael S.
- Fischer, Eric S.
- ORCID
- Venue
- Nature Chemical Biology
- Published
- 2021-05-25
- Volume
- 17
- Issue
- 6
- Pages
- 711-717
- ISSN
- 1552-4450 (ISSN-L)
- 1552-4469
- PMC
- PMC8162940
- OpenAlex
- W3165703330
- Cited by
- 124
- References
- 43
- Citations by year
- 2026: 20 · 2025: 36 · 2024: 26 · 2023: 27 · 2022: 15
- FWCI
- 5.41
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature Chemical Biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Damon Runyon Cancer Research Foundation (DRG-2270-16, DRR-50-18)
- U.S. Department of Health & Human Services | NIH | National Cancer Institute (R01CA214608)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
- Keywords
- Helios, regulatory T cells, small-molecule degraders, cereblon, E3 ubiquitin ligase, zinc finger transcription factor, Treg suppressive function, structure-based drug design
- MeSH terms
- Animals, Cell Line, DNA-Binding Proteins, Humans, Models, Molecular, Mutation, Substrate Specificity, Transcription Factors, Molecular Structure, Jurkat Cells, Ubiquitin-Protein Ligases, Adaptor Proteins, Signal Transducing, T-Lymphocytes, Regulatory, Mice, Ikaros Transcription Factor, Small Molecule Libraries
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-10.
Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-10.
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