The 24 genes that respond to BOTH menin inhibitors in RS4;11 are led by the ATF4 stress programme - CHAC1 -5.11, STC2 -4.40, TRIB3 -3.10 - and CHAC1 and TRIB3 are named zero times in this graph
also called ATF4 stress programme · CHAC1 TRIB3 menin inhibitor · compound-independent menin signature · integrated stress response KMT2A
Comparing a weak and a strong menin-blocking drug in the same leukaemia cells, only twenty-four genes respond to both, and most of them belong to one stress-response programme the cell runs when it is short of nutrients. Two of the strongest are genes this project has never once mentioned, and the single existing paper linking menin to that programme is about a different fusion cancer entirely.
The same thing, for a scientist
Naming the 24 concordant genes from the two-compound intersection turns a methodological result into a biological one. **Down in both (16):** CHAC1 (MI-2-2 -1.38 / VTP-50469 **-5.11**), STC2 (-1.43 / **-4.40**), RGS16 (-1.32 / -3.54), EMID1, GDAP1L1, FAM69C, TRIB3 (-1.23 / **-3.10**), TTC28, SERPINE2, FAM129A, ATF3 (-1.16 / -1.84), SESN2 (-1.32 / -1.42), SOCS2, TSC22D3, RHEBL1, KCNK12. **Up in both (8):** CD84 (+1.11 / **+4.18**), SGK1, RGS2, ZNF503, KIAA1598, CASZ1, KIAA0754, **IKZF3**. **The down list is an ATF4 signature.** CHAC1 is a direct proapoptotic component of the unfolded protein response downstream of the ATF4-ATF3-CHOP cascade (PMID 19109178); TRIB3 is induced via the ATF4-CHOP pathway (PMID 15775988); ATF3 is in that cascade by name; SESN2 and STC2 are ATF4/ISR-responsive. And **ATF5**, an ATF4-family member, falls on all four array probes. **CORRECTED CYCLE 58: the figures -3.02/-2.89/-2.77 are SEM, not RS4;11**, which gives **-1.09/-1.12/-0.74** on the same three probes; the sentence sat inside an RS4;11-scoped argument and read as RS4;11. What survives is stronger than the number it replaces: all four ATF5 probes fall in **both** t(4;11) lines and are flat-to-positive in REH, across baselines 8.07-12.53 log2 RMA. **ALSO CORRECTED: the core is not the 'menin-inhibitor' response.** The same deposited series carries an **EPZ5676 (DOT1L inhibitor)** arm that reproduces this entire collapse at RS4;11 day 7 to within **0.02-0.24 log2** - CHAC1 -5.11/-5.03, STC2 -4.40/-4.16, TRIB3 -3.10/-3.07, ATF5 -2.43/-2.36, ATF4 -0.86/-0.87 - while the menin-dependent arm discriminates by 1.0-3.8 log2 (MEIS1 -5.25/-1.50, MEF2C -4.35/-1.71). So the compound-independent core is **loss of ATF4 output as a FUSION-programme readout, not a menin-interface readout**, and the ISR signature cannot serve as a pharmacodynamic marker of menin engagement. See `hyp-the-isr-collapse-is-a-fusion-programme-readout-not-a-menin-interface-readout`. **This graph is blind to it.** `CHAC1` and `TRIB3` are each named **zero times** across 1,993 entries. There is no `gene-atf4`, `gene-atf3`, `gene-atf5`, `gene-chac1`, `gene-trib3` or `gene-sesn2`. ATF4 itself is named 22 times and "integrated stress response" 16, so the concept is present and the components are not. **And the precedent is already in the graph, in another fusion family.** PMID 33741715 - the ONLY paper PubMed returns for `menin[tiab] AND ATF4[tiab]`, and cited here twice - reports that **EWS-FLI1 and menin converge to regulate ATF4 activity** in Ewing sarcoma, with menin driving serine biosynthesis through it. This graph cites that paper in a sarcoma-risk context and has never connected it to a KMT2A-fusion measurement. PHGDH is named 35 times here; PSAT1 zero. **The discordant genes tell a second story.** Eight genes move oppositely, and the two largest are **ABCG1 (+3.48 under MI-2-2, -1.31 under VTP)** and **ABCA1 (+2.73, -3.96)** - the canonical LXR cholesterol-efflux pair, with `gene-nr1h2` and `gene-nr1h3` already in this graph. A lipophilic tool compound at 10 uM inducing ABCA1/ABCG1 is what an off-target sterol perturbation looks like, and it is a candidate explanation for part of the MI-2-2-specific response. **AND THE SIGNATURE TRACKS THE FUSION.** **CORRECTED CYCLE 58: 23 of the 24, not all 24.** STC2's REH baseline is 4.89 log2 RMA, at background, so its flat response in REH is absence of signal rather than evidence of fusion-dependence. The remaining 23 have a usable probe in **REH**, the KMT2A-wild-type line assayed in the same batch with the same compound at the same dose, and **none of them moves by 1 log2 there** - only TRIB3 (-0.68) exceeds 0.5. They are not missing from REH: TSC22D3 sits at 11.37, TRIB3 at 10.31, SESN2 at 9.99, CHAC1 at 8.95. Expressed in the wild-type line, unresponsive in it, and down 1.0-5.1 log2 in both KMT2A::AFF1 lines. **This refutes the prediction this entry derived from the Ewing paper** - that if menin were the shared node, inhibition should lower ATF4 output without a fusion - with the caveat that MI-2-2's whole-transcriptome effect in REH was small (80 probes), so a null there is weaker than a null in a responsive line. The decisive test, VTP-50469 in a wild-type line, does not exist: GSE127506 contains no wild-type line at all.
| script | analyses/was-the-weak-inhibitor-doing-the-same-thing-weakly/run.py |
|---|---|
| results | analyses/was-the-weak-inhibitor-doing-the-same-thing-weakly/results.json |
| derivation | The 24 concordant and 8 discordant genes were extracted from the same universe of 9,595 genes expressed above floor in both DMSO arms of RS4;11, by importing the analysis module and re-running its set logic rather than by re-deriving thresholds. |
| the atf4 identification | CHAC1 downstream of ATF4-ATF3-CHOP (PMID 19109178, Mungrue, J Immunol 2009); TRIB3 induced via ATF4-CHOP (PMID 15775988, Ohoka, EMBO J 2005); ATF3 is a named member of that cascade; SESN2 and STC2 are ISR-responsive. ATF5, an ATF4-family member, is the most reproducible repression in the companion dataset. |
| the precedent in another fusion family | PMID 33741715 (Jimenez JA, Apfelbaum AA, Hawkins AG, et al., 'EWS-FLI1 and Menin Converge to Regulate ATF4 Activity in Ewing Sarcoma', Mol Cancer Res, doi 10.1158/1541-7786.MCR-20-0679) is the ONLY record for menin[tiab] AND ATF4[tiab] - count 1, parsed translation. It is already cited in two entries here, both in a sarcoma-risk context, and never connected to a KMT2A-fusion measurement. |
| citation year discrepancy | This graph's stored citation for PMID 33741715 says 2021; efetch returns 'Molecular cancer research : MCR 2022'. Most likely an epub-2021 / issue-2022 split rather than an error, and it is recorded here rather than silently changed in the two entries that carry it. |
| shown for fusion | true |
| absence class | measured-not-analysed |
concordant down 16
{
"CHAC1": "MI-2-2 -1.38 / VTP -5.11 (array DMSO 8.32, rna DMSO 6.35)",
"STC2": "-1.43 / -4.40 (8.62, 5.02)",
"RGS16": "-1.32 / -3.54 (8.01, 7.45)",
"EMID1": "-1.40 / -3.20",
"GDAP1L1": "-1.41 / -3.10",
"FAM69C": "-1.38 / -3.10",
"TRIB3": "-1.23 / -3.10 (8.78, 7.57)",
"TTC28": "-1.08 / -2.86",
"SERPINE2": "-1.85 / -1.96",
"FAM129A": "-1.55 / -1.82",
"ATF3": "-1.16 / -1.84 (8.52, 4.74)",
"SESN2": "-1.32 / -1.42 (9.43, 7.33)",
"SOCS2": "-1.65 / -1.06 (11.16, 5.61)",
"TSC22D3": "-1.04 / -1.66 (11.05, 7.70)",
"RHEBL1": "-1.12 / -1.35",
"KCNK12": "-1.07 / -1.08"
}concordant up 8
{
"CD84": "MI-2-2 +1.11 / VTP +4.18",
"SGK1": "+1.00 / +1.99",
"RGS2": "+1.55 / +1.84",
"ZNF503": "+1.08 / +1.60",
"KIAA1598": "+1.65 / +1.35",
"CASZ1": "+1.21 / +1.39",
"KIAA0754": "+1.11 / +1.41",
"IKZF3": "+1.08 / +1.34"
}discordant 8
{
"ABCG1": "MI-2-2 +3.48 / VTP -1.31",
"ABCA1": "+2.73 / -3.96",
"NCF1": "+1.03 / -3.96",
"FLT1": "+1.15 / -3.48",
"PIGZ": "-1.10 / +2.04",
"BLVRA": "-1.26 / +2.02",
"PAQR8": "-1.07 / +1.93",
"PMP22": "-1.05 / +1.45"
}what this graph holds and does not
{
"CHAC1 mentions": 0,
"TRIB3 mentions": 0,
"ATF4 mentions": 22,
"integrated stress response mentions": 16,
"PHGDH mentions": 35,
"PSAT1 mentions": 0,
"entries that exist": "gene-nr1h2 and gene-nr1h3 (LXRbeta and LXRalpha), fusion-ewsr1-fli1",
"entries that do not": "gene-atf4, gene-atf3, gene-atf5, gene-chac1, gene-trib3, gene-sesn2, gene-abca1, gene-abcg1, gene-ikzf3, gene-sgk1, gene-socs2",
"a_dangling_edge_i_wrote": "I wrote an edge to gene-socs2 from this entry without checking the node existed. It does not, and validate-graph.mjs caught it as a dangling target. SOCS2 falls under both compounds (-1.65 / -1.06) and not in the wild-type line (-0.34), so it belongs on the named-unwritten list rather than on an edge. Same failure as the agent this project caught last cycle writing an edge to gene-srpk2 from recall."
}the wild type discriminator
{
"result": "0 of 24 concordant genes move >=1 log2 in REH under MI-2-2; 1 (TRIB3, -0.68) exceeds 0.5 in the same direction.",
"they_are_expressed_there": {
"TSC22D3": 11.37,
"FAM129A": 11.22,
"SGK1": 10.89,
"TRIB3": 10.31,
"SESN2": 9.99,
"TTC28": 9.49,
"FAM69C": 9.9,
"CHAC1": 8.95,
"ATF3": 8.75,
"SOCS2": 9.77
},
"reh_values": {
"ATF3": 0.055,
"CASZ1": 0.056,
"CD84": 0.263,
"CHAC1": 0.494,
"EMID1": 0.114,
"FAM129A": -0.243,
"FAM69C": 0.052,
"GDAP1L1": -0.289,
"IKZF3": 0.088,
"KCNK12": -0.175,
"KIAA0754": 0.065,
"KIAA1598": -0.228,
"RGS16": -0.196,
"RGS2": 0.123,
"RHEBL1": -0.262,
"SERPINE2": -0.183,
"SESN2": -0.042,
"SGK1": 0.032,
"SOCS2": -0.338,
"STC2": -0.332,
"TRIB3": -0.678,
"TSC22D3": -0.063,
"TTC28": 0.005,
"ZNF503": 0.13
},
"reading": "The genes are present in the wild-type line and unresponsive in it. That makes the signature fusion-tracking, and it refutes the prediction derived from the Ewing menin-ATF4 paper."
}CORRECTED CYCLE 58
{
"who": "four defects found by Q106 re-running the same cached matrices; the fifth verified by the king",
"1_atf5_line_mislabel": {
"was": "-3.02/-2.89/-2.77 on three MI-2-2 probes, presented inside an RS4;11 argument",
"is": "those are SEM; RS4;11 gives -1.09/-1.12/-0.74",
"what_survives": "all FOUR ATF5 probes fall in BOTH t(4;11) lines, flat-to-positive in REH"
},
"2_stc2_has_no_usable_reh_probe": {
"was": "all 24 genes have a usable probe in REH",
"is": "23 of 24. STC2's REH baseline is 4.89 log2 RMA - background - so its flat REH response is absence of signal."
},
"3_the_frame_was_wrong": {
"was": "the compound-independent core of the MENIN-INHIBITOR response",
"is": "compound-independent across TWO DIFFERENT TARGETS. The EPZ5676 DOT1L arm in the same series reproduces the ISR collapse to within 0.02-0.24 log2 while the menin arm discriminates by 1.0-3.8 log2 on MEIS1/MEF2C/PBX3/PHGDH. The assay HAS the resolution to detect menin specificity and detects none on the ISR arm.",
"consequence": "the ISR/ATF4 signature cannot be a pharmacodynamic marker of menin engagement - either drug produces it - and a menin/DOT1L combination buys nothing on this axis. Fusion-dependence survives: nothing moves in REH."
},
"4_two_floor_collisions_in_the_concordant_down_list": {
"GDAP1L1": "-3.10 with the DMSO baseline at exactly 3.10 and the treated arm at zero counts",
"KCNK12": "-1.08 with baseline 1.08",
"so": "these are detection-floor artefacts, not measured fold changes, and should not have been listed as concordant hits"
},
"5_the_precedent_is_NOT_in_another_fusion_family_entirely": {
"was": "the precedent is in another fusion family entirely",
"is": "PMID 33741715 is primarily Ewing sarcoma but its own abstract states that menin inhibition 'similarly leads to loss of ATF4 and the ATF4-dependent transcriptional signature in MLL-rearranged B-cell acute lymphoblastic leukemia', and that menin-dependent regulation of ATF4 is 'transcriptional and post-transcriptional'. A published menin-inhibition ATF4 result in the INDEX DISEASE CLASS already existed.",
"and_it_anticipated_the_mechanism": "the post-transcriptional clause anticipates cycle 58's finding that ATF4's own transcript does not move",
"and_it_converges_with_this_graph_elsewhere": "that paper runs menin -> ATF4 -> SERINE biosynthesis, which is the same axis as this project's PHGDH/PSAT1/ASNS findings from cycles 55-57",
"verified_by": "king, abstract re-fetched 2026-09-22"
}
}- This is a list of genes from two cell-line datasets, not a therapeutic claim. Nothing here says a menin inhibitor should or should not be given to anyone, and the ATF4 identification is of a signature by its targets - no ATF4 protein or activity was measured in either dataset.
- The ISR reading has a mundane alternative that cannot be excluded here: a drug that slows growth lowers translational demand, so stress-response output can fall as a consequence of the phenotype rather than as part of the mechanism. Any citation of this entry should carry that.
- If the ATF4 axis were pursued therapeutically, note the published hazard: loss of ATF4 causes aging-like functional attrition of adult haematopoietic stem cells with defective repopulation and self-renewal (PMID 34936448, Sun et al., Science Advances 2023, fetched and verified). A programme whose loss damages normal HSCs is a poor candidate for further suppression in an infant, and this entry should not be used to propose one.
- Twenty-four genes with ~25% expected to be coincidence is not a validated signature. Individual genes on the list are not established as menin-dependent; only the set carries the signal.
- Is ATF4 protein or activity actually reduced by menin inhibition in a KMT2A-rearranged cell? Nothing here measures ATF4 - only its targets - and the inversion (ISR output falling under an effective drug) makes the direct measurement worth doing rather than assuming.
- ANSWERED the same cycle, and it went against the prediction. All 24 concordant genes were extracted from REH, the KMT2A-wild-type line in the same GSE63664 batch under the same MI-2-2 exposure: 0 of 24 move by 1 log2 and only TRIB3 (-0.68) exceeds 0.5, while their REH baselines are high (CHAC1 8.95, TRIB3 10.31, TSC22D3 11.37). The signature tracks the fusion. The prediction from the Ewing paper - that menin inhibition should lower ATF4 output without a fusion because the shared node is menin - is not supported, with the caveat that MI-2-2's whole-transcriptome effect in REH was small (80 probes). See E4. The decisive test, VTP-50469 in a wild-type line, does not exist in GSE127506.
- Is the ISR collapse a cause or a consequence? A drug that slows growth lowers translational demand and ISR output. A growth-matched control arm, or an early timepoint before the phenotype, would separate them; GSE127506's day-2 arm is the natural place to look and MEIS1 is already -2.59 there.
- CHAC1 and TRIB3 have no entry and are named zero times in 1,993 entries. So do ATF4, ATF3, ATF5, SESN2, ABCA1, ABCG1, IKZF3 and SGK1. IKZF3 is the most interesting of those for this disease specifically, given that IKZF1 deletion is the recurrent cooperating lesion - and IKZF3 goes UP under both compounds.
- Is the ABCA1/ABCG1 induction under MI-2-2 an LXR effect, and does it explain part of the MI-2-2-only response? gene-nr1h2 and gene-nr1h3 exist in this graph. A lipophilic compound at 10 uM inducing both canonical LXR efflux targets while the potent compound at 330 nM represses them is the cleanest off-target signal either dataset offers.
names the 24 concordant genes that analysis only counted, and adds the wild-type discriminator: 0 of 24 respond in REH
- analogous to →EWSR1::FLI1 (EWS-FLI1), t(11;22)(q24;q12)
the only menin-ATF4 paper in PubMed reports EWS-FLI1 and menin converging on ATF4 in Ewing sarcoma; this graph cited it for sarcoma risk and never connected it to a KMT2A measurement
ABCA1 and ABCG1 rise under MI-2-2 at 10 uM and FALL under VTP-50469 at 330 nM -- a candidate LXR/sterol off-target signature for the weak compound
the second LXR receptor already in this graph; the ABCA1/ABCG1 discordance is the cleanest off-target signal either dataset offers
grew out of that entry ATF4 identification: naming the 24 concordant genes pointed at ATF4, and the serine/ASNS arm was then checked
partially refutes the ATF4/ISR framing: of the five genes that identified the signature, TRIB3 and ATF3 reverse sign in an independent line and STC2 is undetected; CHAC1 and ATF5 hold
the ISR arm of that entry's 24-gene signature is reproduced to within 0.02-0.24 log2 by EPZ5676, an arm that entry never reported, so it is not a menin-interface readout; and the ATF5 triple it quotes is SEM, not RS4;11