Figures and data in A switch in transcription and cell fate governs the onset of an epigenetically-deregulated tumor in Drosophila

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Joana Torres

Remo Monti

Ariane L Moore

Makiko Seimiya

Yanrui Jiang

Niko Beerenwinkel

Christian Beisel

Jorge V Beira

Renato Paro

(2018)
A switch in transcription and cell fate governs the onset of an epigenetically-deregulated tumor in Drosophila

eLife 7:e32697.

https://doi.org/10.7554/eLife.32697
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Additional files

13 figures, 1 table and 1 additional file

Figures

Figure 1 with 4 supplements

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Gene expression analysis of *ph⁵⁰⁵*-tumor cells reveals loss of cell identity and acquisition of an embryonic-TF signature.

RNA-sequencing was performed in FACS-sorted *ph⁵⁰⁵* tissue samples. Gene expression in tumor cells (GFP⁺) was compared with surrounding wild-type cells (GFP^-) from the same pool of eye-antennal …
https://doi.org/10.7554/eLife.32697.003

Figure 1—source data 1 List of differentially expressed genes.: https://doi.org/10.7554/eLife.32697.008
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Figure 1—source data 2 Gene Ontology (GO) analysis.: https://doi.org/10.7554/eLife.32697.009
Download elife-32697-fig1-data2-v1.xlsx
Figure 1—source data 3 Information relative to samples used for clustering (83 samples).: https://doi.org/10.7554/eLife.32697.010
Download elife-32697-fig1-data3-v1.xlsx
Figure 1—source data 4 List of genes used for hierarchical clustering analysis.: https://doi.org/10.7554/eLife.32697.011
Download elife-32697-fig1-data4-v1.xlsx
Figure 1—source data 5 List of Genotypes.: https://doi.org/10.7554/eLife.32697.012
Download elife-32697-fig1-data5-v1.xlsx

Figure 1—figure supplement 1

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Neoplastic growth of ph mutant clones in eye-antennal imaginal discs can be rescued by wt ph co-expression.

Mitotic clones were generated by using eyFlp/FRT and MARCM systems, allowing for generation of GFP-labeled clones within eye-antennal imaginal discs. Ph expression in FRT19A control (**A–A’’’**) and in p…
https://doi.org/10.7554/eLife.32697.004

Figure 1—figure supplement 2

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Gene expression analysis of *ph⁵⁰⁵*-tumor cells reveals loss of cell identity and acquisition of an embryonic-TF signature.

Heatmap of normalized expression levels of 1337 differentially expressed genes across tumor and control samples (A). A total of 1337 genes are differentially expressed in tumor samples in comparison …
https://doi.org/10.7554/eLife.32697.005

Figure 1—figure supplement 3

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*ph⁵⁰⁵*-tumor cells do not express differentiation markers.

Normal pattern (A) expression of ELAV retinal cell marker is interrupted in *ph⁵⁰⁵* clones (B). Eyes absent (Eya) and Homothorax (Hth) protein expression in FRT neutral (**C, E**) and *ph⁵⁰⁵* (**D, F**) clones. …
https://doi.org/10.7554/eLife.32697.006

Figure 1—figure supplement 4

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*ph⁵⁰⁵*-tumor cells ectopically express embryonic transcription factors.

Embryonic TF Abdominal-B (Abd-B) and Caudal (Cad) are ectopically expressed in *ph⁵⁰⁵* clones (**B–D**) in comparison to FRT19A control tissues (**A–C**). Scale bar corresponds to 100 µm. All microscope …
https://doi.org/10.7554/eLife.32697.007

Figure 2 with 2 supplements

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KD of embryonic TFs in *ph⁵⁰⁵*-tumors can increase the viability of the flies and reduce tumor volume.

Eclosion rates (%) for FRT19A control, *ph⁵⁰⁵* and for specific KD in *ph⁵⁰⁵* background (*ph⁵⁰⁵* +UAS RNAi). Fly stocks carrying RNAi constructs were used for the KD of 24 TFs upregulated in *ph⁵⁰⁵*-tumor …
https://doi.org/10.7554/eLife.32697.013

Figure 2—figure supplement 1

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KD of embryonic TFs in *ph⁵⁰⁵*-tumors can increase the viability of the flies and reduce tumor volume.

Example of eye-antennal imaginal disc carrying *ph⁵⁰⁵* cells used for quantification of tumor volume (A). Average tumor volume (µm³) (B), tissue volume (µm³) (C) and number of clones *per* tissue (D) …
https://doi.org/10.7554/eLife.32697.014

Figure 2—figure supplement 2

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KD of embryonic TFs in *ph⁵⁰⁵*-tumors can increase the viability of the flies and reduce tumor volume.

Representative adult eyes of 4 different genotypes: FRT19A neutral clones (A), FRT19A, kni-KD (B), *ph⁵⁰⁵* (C) and *ph⁵⁰⁵*, kni-KD (D). ELAV protein expression in *ph⁵⁰⁵*, kni-KD tissues (E). Higher …
https://doi.org/10.7554/eLife.32697.015

Figure 3 with 1 supplement

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kni-KD in *ph⁵⁰⁵*-tumors has anti-oncogenic effects Percentage of *ph⁵⁰⁵* tumor-bearing hosts over time ranges from 40–60% (A).

Female host shows abdominal tumor growth one week after transplantation (**B–B’**). Percentage of *ph⁵⁰⁵*, kni-KD tumor-bearing hosts is shown over time and ranges from 0% to 3% (C). One single host with …
https://doi.org/10.7554/eLife.32697.016

Figure 3—figure supplement 1

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kni-KD in *ph⁵⁰⁵*-tumors has anti-oncogenic effects.

Levels of apoptosis (Dcp-1) are shown for FRT19A, kni-KD (**A–A’**) and FRT19A, kni-KD, UAS-p35 (**B–B’**). Higher magnification insets are shown in A’ and B’. Eclosion rates for the same conditions (C). …
https://doi.org/10.7554/eLife.32697.017

Figure 4

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Knirps misexpression induces cyst formation in imaginal discs and is sufficient to drive tumorigenesis.

Ectopic expression of *knirps* in eye-antennal imaginal discs shows a distinct pattern of clones (**A–A’**). Eclosion rate (B) for FRT19A and FRT19A, UAS-Kni. Ectopic expression of *kni* in these particular …
https://doi.org/10.7554/eLife.32697.018

Figure 5 with 1 supplement

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Knirps misexpression induces JAK/STAT activation and compromises differentiation.

Activity of JAK/STAT, JNK and Notch signaling pathways in the context of kni-ectopic expression in eye-antennal discs was assessed by evaluating the expression of the respective activity reporters: …
https://doi.org/10.7554/eLife.32697.019

Figure 5—figure supplement 1

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Knirps misexpression induces JAK/STAT activation and compromises differentiation.

Disruption of Armadillo protein expression is observed in UAS-Kni expressing clones (A). ELAV protein expression is interrupted in *ph⁵⁰⁵*, dome.∆CYT (blockage of JAK/STAT pathway in *ph⁵⁰⁵* background) …
https://doi.org/10.7554/eLife.32697.020

Figure 6 with 2 supplements

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Overexpression of pro-neural TF in *ph⁵⁰⁵*-clones suppresses tumor phenotype.

Forcing differentiation of *ph⁵⁰⁵* cells by ectopically expressing pro-neural TF *atonal* leads to a reduction of tumor volume (**A–B**). Number of tissues analyzed: *ph⁵⁰⁵* N = 50; *ph⁵⁰⁵*, UAS-ato N = 23. On …
https://doi.org/10.7554/eLife.32697.021

Figure 6—figure supplement 1

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Overexpression of pro-neural TF in *ph⁵⁰⁵*-clones suppresses tumor phenotype.

Ectopic expression of pro-neural TF *atonal* in FRT19A neutral clones (A). Average tumor volume (µm³) (B), tissue volume (µm³) (C) and number of clones *per* tissue (D) for the ectopic expression of *ato* …
https://doi.org/10.7554/eLife.32697.022

Figure 6—figure supplement 2

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Overexpression of pro-neural TF in *ph⁵⁰⁵*-clones prevents cells from proliferating.

Proliferation levels (phospho-histone H3, pH3) upon overexpressing *ato* in control tissues (**A–B**). Quantitative analysis of pH3+ cell numbers in FRT19A; FRT19A, UAS-ato; *ph505* and *ph505*, UAS-ato (C). …
https://doi.org/10.7554/eLife.32697.023

Author response image 1

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Eye-antennal imaginal discs from different developmental stages carrying ph⁵⁰⁵-clones (A-C).

Mutant cells are GFP labeled and present in late first instar EA discs (A). Pictures were taken with the same objective in a wide-field microscope (scale bar = 100μm). EA, eye-antennal imaginal …
https://doi.org/10.7554/eLife.32697.026

Author response image 2

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Expression of ELAV (magenta) in developing eye-antennal imaginal discs (EA), 96h and 120h after egg laying (AEL).

White arrows, morphogenetic furrow. DAPI, cyan; magenta, ELAV staining. Scale bar 100μm.

https://doi.org/10.7554/eLife.32697.027

Author response image 3

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Expression of ELAV in mitotic clones generated 96h AEL.

Heat shock (HS) experiments were performed at 96h AEL (90min, 37ºC). Control experiments were done with FRT19A neutral clones (left side, GFP clones), where clones are very small at 6 days AEL, and …
https://doi.org/10.7554/eLife.32697.028

Author response image 4

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Differential Expression analysis – comparison of ph⁵⁰⁵-tumor cells (GFP+) with wildtype EA discs.

(A) Venn diagram showing the overlap (1088 DE genes) between the previous analysis (DE vs. GFP-) and the new analysis (DE vs. EA). (B) UpSet intersection diagram shows detail information about …
https://doi.org/10.7554/eLife.32697.029

Author response image 5

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Generation of ph⁵⁰⁵ mutant clones in wing imaginal discs.

(A) Control clones generated with FRT19A blank and UbxFlp present notchy-shape (A’) and do not express Caudal (A’’). (B) ph⁵⁰⁵ clones generated in wing imaginal discs, similarly to what was observed …
https://doi.org/10.7554/eLife.32697.030

Author response image 6

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JAK/STAT and JNK (MMP1) pathway activation in *ph⁵⁰⁵* clones in wing imaginal discs.

(A) Expression of the 10x-STAT reporter in wild-type wing imaginal disc. (B) ph⁵⁰⁵ clones generated in wing imaginal discs show ectopic expression of the reporter inside the pouch (B’-B’’’). (C) …
https://doi.org/10.7554/eLife.32697.031

Author response image 7

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Quantitative analysis of tissue volume occupied by ELAV+ cells.

Number of tissues used for quantification analysis (using Imaris): FRT19A N=12; ph⁵⁰⁵ N=19). Trend of increase in absolute volume of ELAV+ inside tissues carrying ph⁵⁰⁵-tumor clones is observed, in …
https://doi.org/10.7554/eLife.32697.032

Tables

Key resources table

Reagent type (species) or resource	Designation	Source or reference	Identifiers	Additional information
genetic reagent (D. melanogaster)	OregonR (host flies)	Bloomington Drosophila Stock Center	BL25211
genetic reagent (D. melanogaster)	w[1118] (host flies)	Bloomington Drosophila Stock Center	BL5905
genetic reagent (D. melanogaster)	yw, FRT19A	Bloomington Drosophila Stock Center	BL1744
genetic reagent (D. melanogaster)	FRT19A, ph505/FM7 act-GFP	A.-M. Martinez	ph505
genetic reagent (D. melanogaster)	w, tubGal80, FRT19A; eyFlp5, Act5C > y +> Gal4, UAS-GFP.S56T	T. Xu	19A Tester
genetic reagent (D. melanogaster)	P{w[+mC]=tubP-GAL80}LL1, w* P{ry[+t7.2]=ey FLP.N}2P{ry[+t7.2]=neoFRT}19A	Bloomington Drosophila Stock Center	BL42717
genetic reagent (D. melanogaster)	w[1118]; P{w[+mC]=GAL4-Act5C(FRT.CD2).P}S, P{w[+mC]=UAS RFP.W}3/TM3	Bloomington Drosophila Stock Center	BL30558
genetic reagent (D. melanogaster)	w; UAS-dome∆CYT	J. Hombría	dome∆CYT
genetic reagent (D. melanogaster)	UAS-ph L7 (#3)	F. Maschat	UAS-ph
genetic reagent (D. melanogaster)	w; UAS-p35	S. Kurata	UAS-p35
genetic reagent (D. melanogaster)	UAS-Kni/Cyo	M. Affolter	UAS-Kni
genetic reagent (D. melanogaster)	UAS-ato (#2)	G. Mardon	UAS-ato
genetic reagent (D. melanogaster)	w1118;; P{NRE-EGFP.S}1	Bloomington Drosophila Stock Center	BL30728 (Notch reporter, NRE-GFP on #3)
genetic reagent (D. melanogaster)	w;;TRE-DsRed	Chatterjee & Bohmann 2012	JNK reporter
genetic reagent (D. melanogaster)	w[1118]; P{w[+mC]=10XStat92E-GFP}2	Bloomington Drosophila Stock Center	BL26198 (JAK/STAT reporter)
genetic reagent (D. melanogaster)	RNAi of Kni: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMS01184}attP2	Bloomington Drosophila Stock Center	Trip BL34705 (kni-KD)
genetic reagent (D. melanogaster)	RNAi of Kni: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF02544}attP2	Bloomington Drosophila Stock Center	Trip BL27259 (kni-KD (2))
genetic reagent (D. melanogaster)	RNAi of Abd-A: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF03167}attP2	Bloomington Drosophila Stock Center	Trip BL28739 (Abd-A-KD)
genetic reagent (D. melanogaster)	RNAi of lms: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMS02709}attP2/TM3, Sb(Abate-Shen, 2002)	Bloomington Drosophila Stock Center	Trip BL43995 (lms-KD)
genetic reagent (D. melanogaster)	RNAi of gsc: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMC02397}attP2	Bloomington Drosophila Stock Center	Trip BL50894 (gsc-KD)
genetic reagent (D. melanogaster)	RNAi of cad: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMC04863}attP40	Bloomington Drosophila Stock Center	Trip BL57546 (cad-KD)
genetic reagent (D. melanogaster)	RNAi of bgcn: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP .GL00596}attP40	Bloomington Drosophila Stock Center	Trip BL36636 (bgcn-KD)
genetic reagent (D. melanogaster)	RNAi of Sox100b: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.GLV21021}attP2	Bloomington Drosophila Stock Center	Trip BL35656 (Sox100b-KD)
genetic reagent (D. melanogaster)	RNAi of btd: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF03389}attP2	Bloomington Drosophila Stock Center	Trip BL29453 (btd-KD)
genetic reagent (D. melanogaster)	RNAi of eve: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF03161}attP2	Bloomington Drosophila Stock Center	Trip BL28734 (eve-KD)
genetic reagent (D. melanogaster)	RNAi of tin: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMC03064}attP2	Bloomington Drosophila Stock Center	Trip BL50663 (tin-KD)
genetic reagent (D. melanogaster)	RNAi of Dr: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMC03402}attP2	Bloomington Drosophila Stock Center	Trip BL51830 (Dr-KD)
genetic reagent (D. melanogaster)	RNAi of nub: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMC03992}attP2	Bloomington Drosophila Stock Center	Trip BL55305 (nub-KD)
genetic reagent (D. melanogaster)	RNAi of fkh: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMS01103}attP2	Bloomington Drosophila Stock Center	Trip BL33760 (fkh-KD)
genetic reagent (D. melanogaster)	RNAi of Abd-B: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF02309}attP2	Bloomington Drosophila Stock Center	Trip BL26746 (Abd-B-KD)
genetic reagent (D. melanogaster)	RNAi of pb: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMC03065}attP2	Bloomington Drosophila Stock Center	Trip BL50664 (pb-KD)
genetic reagent (D. melanogaster)	RNAi of grn: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMS01085}attP2	Bloomington Drosophila Stock Center	Trip BL33746 (grn-KD)
genetic reagent (D. melanogaster)	RNAi of odd: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMS01315}attP2/TM3, Sb(Abate-Shen, 2002)	Bloomington Drosophila Stock Center	Trip BL34328 (odd-KD)
genetic reagent (D. melanogaster)	RNAi of croc: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMS01122}attP2	Bloomington Drosophila Stock Center	Trip BL34647 (croc-KD)
genetic reagent (D. melanogaster)	RNAi of drm: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMJ02120}attP40	Bloomington Drosophila Stock Center	Trip BL42548 (drm-KD)
genetic reagent (D. melanogaster)	RNAi of Doc2: y(Abate-Shen, 2002) sc[*] v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.HMS02804}attP2	Bloomington Drosophila Stock Center	Trip BL44087 (Doc2-KD)
genetic reagent (D. melanogaster)	RNAi of Doc3: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF02223}attP2	Bloomington Drosophila Stock Center	Trip BL31932 (Doc3-KD)
genetic reagent (D. melanogaster)	RNAi of vvl: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF02126}attP2	Bloomington Drosophila Stock Center	Trip BL26228 (vvl-KD)
genetic reagent (D. melanogaster)	RNAi of Doc1: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF02222}attP2	Bloomington Drosophila Stock Center	Trip BL31931 (Doc1-KD)
genetic reagent (D. melanogaster)	RNAi of Kr: y(Abate-Shen, 2002) v(Abate-Shen, 2002); P{y[+t7.7] v[+t1.8]=TRiP.JF02745}attP2	Bloomington Drosophila Stock Center	Trip BL27666 (Kr-KD)
antibody	Ph (rabbit)	R. Paro Lab	N/A	(1:100)
antibody	Arm (mouse)	DSHB	N27A1	(1:5)
antibody	MMP1 (mouse)	DSHB	5H7B11	(1:300)
antibody	ELAV (rat)	DSHB	7E8A10	(1:30)
antibody	Eya (mouse)	DSHB	eya10H6	(1:500)
antibody	Hth (goat)	H. Sun	dG20, Santa Cruz	(1:100)
antibody	Eve (mouse)	DSHB	Eve 3C10	(1:100)
antibody	Cad (rabbit)	P. Macdonald Lab	#1	(1:500)
antibody	Abd-B (mouse)	DSHB	N/A	(1:10)
antibody	Dcp-1 (rabbit)	Cell Signaling	9578S	(1:200)
antibody	Phalloidin Alexa 633	Life Technologies	A22284	(1:400)
antibody	phospho-Histone H3, Ser 10 (pH3, rabbit)	Millipore	06–570	(1:200)
antibody	Alexa 568- or 594 secondaries	Life Technologies	A-11036, A11031, A-11077, A-11058	(1:500)

commercial assay or kit	PicoPure RNA Isolation Kit	Thermo Fisher	KIT0204
commercial assay or kit	RNase-Free DNase Set	Qiagen	#79254
commercial assay or kit	Quant-iT RiboGreen RNA Assay Kit	Thermo Fisher	R11490
chemical compound, drug	Collagenase	Sigma	C1639

software, algorithm	iRegulon	Janky et al. (2014)	http://iregulon.aertslab.org
software, algorithm	Trimmomatic	Bolger et al. (2014)	http://www.usadellab.org/cms/?page=trimmomatic
software, algorithm	FastQC	FastQC A Quality Control tool for High Throughput Sequence Data (v0.11.2)	www.bioinformatics.babraham.ac.uk/projects/fastqc/
software, algorithm	STAR	Dobin et al. (2013)	https://github.com/alexdobin/STAR
software, algorithm	Picard Tools	Picard tools (version 1.121)	www.broadinstitute.github.io/picard/
software, algorithm	HTSeq	Anders et al. (2015)	http://www-huber.embl.de/HTSeq/doc/overview.html#
software, algorithm	DESeq2	Love et al. (2014)	https://bioconductor.org/packages/release/bioc/html/DESeq2.html
software, algorithm	WEB-based GEne SeT AnaLysis Toolkit	Wang et al., 2013	WebGestalt: www.webgestalt.org
software, algorithm	Ilastik	Sommer, 2011	http://ilastik.org
software, algorithm	Matlab	MATLAB 2016b, The MathWorks Inc., Natick, MA	https://ch.mathworks.com/products/matlab.html
software, algorithm	Imaris	Imaris v 8.4.1 (Build 41809 for x64), Bitplane AG	http://www.bitplane.com/imaris/imaris
software, algorithm	GraphPad Prism 7.0	GraphPad Prism version 7.00 for Windows, GraphPad Software, La Jolla California USA	https://www.graphpad.com/scientific-software/prism/

Additional files

Transparent reporting form: https://doi.org/10.7554/eLife.32697.024
Download elife-32697-transrepform-v1.docx

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Figures

Gene expression analysis of ph⁵⁰⁵-tumor cells reveals loss of cell identity and acquisition of an embryonic-TF signature.

Figure 1—source data 1

Figure 1—source data 2

Figure 1—source data 3

Figure 1—source data 4

Figure 1—source data 5

Neoplastic growth of ph mutant clones in eye-antennal imaginal discs can be rescued by wt ph co-expression.

Gene expression analysis of ph⁵⁰⁵-tumor cells reveals loss of cell identity and acquisition of an embryonic-TF signature.

ph⁵⁰⁵-tumor cells do not express differentiation markers.

ph⁵⁰⁵-tumor cells ectopically express embryonic transcription factors.

KD of embryonic TFs in ph⁵⁰⁵-tumors can increase the viability of the flies and reduce tumor volume.

KD of embryonic TFs in ph⁵⁰⁵-tumors can increase the viability of the flies and reduce tumor volume.

KD of embryonic TFs in ph⁵⁰⁵-tumors can increase the viability of the flies and reduce tumor volume.

kni-KD in ph⁵⁰⁵-tumors has anti-oncogenic effects Percentage of ph⁵⁰⁵ tumor-bearing hosts over time ranges from 40–60% (A).

kni-KD in ph⁵⁰⁵-tumors has anti-oncogenic effects.

Knirps misexpression induces cyst formation in imaginal discs and is sufficient to drive tumorigenesis.

Knirps misexpression induces JAK/STAT activation and compromises differentiation.

Knirps misexpression induces JAK/STAT activation and compromises differentiation.

Overexpression of pro-neural TF in ph⁵⁰⁵-clones suppresses tumor phenotype.

Overexpression of pro-neural TF in ph⁵⁰⁵-clones suppresses tumor phenotype.

Overexpression of pro-neural TF in ph⁵⁰⁵-clones prevents cells from proliferating.

Eye-antennal imaginal discs from different developmental stages carrying ph⁵⁰⁵-clones (A-C).

Expression of ELAV (magenta) in developing eye-antennal imaginal discs (EA), 96h and 120h after egg laying (AEL).

Expression of ELAV in mitotic clones generated 96h AEL.

Differential Expression analysis – comparison of ph⁵⁰⁵-tumor cells (GFP+) with wildtype EA discs.

Generation of ph⁵⁰⁵ mutant clones in wing imaginal discs.

JAK/STAT and JNK (MMP1) pathway activation in ph⁵⁰⁵ clones in wing imaginal discs.

Quantitative analysis of tissue volume occupied by ELAV+ cells.

Tables

Additional files

Transparent reporting form

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Gene expression analysis of ph505-tumor cells reveals loss of cell identity and acquisition of an embryonic-TF signature.

Figure 1—source data 1

Figure 1—source data 2

Figure 1—source data 3

Figure 1—source data 4

Figure 1—source data 5

Neoplastic growth of ph mutant clones in eye-antennal imaginal discs can be rescued by wt ph co-expression.

Gene expression analysis of ph505-tumor cells reveals loss of cell identity and acquisition of an embryonic-TF signature.

ph505-tumor cells do not express differentiation markers.

ph505-tumor cells ectopically express embryonic transcription factors.

KD of embryonic TFs in ph505-tumors can increase the viability of the flies and reduce tumor volume.

KD of embryonic TFs in ph505-tumors can increase the viability of the flies and reduce tumor volume.

KD of embryonic TFs in ph505-tumors can increase the viability of the flies and reduce tumor volume.

kni-KD in ph505-tumors has anti-oncogenic effects Percentage of ph505 tumor-bearing hosts over time ranges from 40–60% (A).

kni-KD in ph505-tumors has anti-oncogenic effects.

Knirps misexpression induces cyst formation in imaginal discs and is sufficient to drive tumorigenesis.

Knirps misexpression induces JAK/STAT activation and compromises differentiation.

Knirps misexpression induces JAK/STAT activation and compromises differentiation.

Overexpression of pro-neural TF in ph505-clones suppresses tumor phenotype.

Overexpression of pro-neural TF in ph505-clones suppresses tumor phenotype.

Overexpression of pro-neural TF in ph505-clones prevents cells from proliferating.

Eye-antennal imaginal discs from different developmental stages carrying ph505-clones (A-C).

Expression of ELAV (magenta) in developing eye-antennal imaginal discs (EA), 96h and 120h after egg laying (AEL).

Expression of ELAV in mitotic clones generated 96h AEL.

Differential Expression analysis – comparison of ph505-tumor cells (GFP+) with wildtype EA discs.

Generation of ph505 mutant clones in wing imaginal discs.

JAK/STAT and JNK (MMP1) pathway activation in ph505 clones in wing imaginal discs.

Quantitative analysis of tissue volume occupied by ELAV+ cells.

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Gene expression analysis of ph⁵⁰⁵-tumor cells reveals loss of cell identity and acquisition of an embryonic-TF signature.

Gene expression analysis of ph⁵⁰⁵-tumor cells reveals loss of cell identity and acquisition of an embryonic-TF signature.

ph⁵⁰⁵-tumor cells do not express differentiation markers.

ph⁵⁰⁵-tumor cells ectopically express embryonic transcription factors.

KD of embryonic TFs in ph⁵⁰⁵-tumors can increase the viability of the flies and reduce tumor volume.

KD of embryonic TFs in ph⁵⁰⁵-tumors can increase the viability of the flies and reduce tumor volume.

KD of embryonic TFs in ph⁵⁰⁵-tumors can increase the viability of the flies and reduce tumor volume.

kni-KD in ph⁵⁰⁵-tumors has anti-oncogenic effects Percentage of ph⁵⁰⁵ tumor-bearing hosts over time ranges from 40–60% (A).

kni-KD in ph⁵⁰⁵-tumors has anti-oncogenic effects.

Overexpression of pro-neural TF in ph⁵⁰⁵-clones suppresses tumor phenotype.

Overexpression of pro-neural TF in ph⁵⁰⁵-clones suppresses tumor phenotype.

Overexpression of pro-neural TF in ph⁵⁰⁵-clones prevents cells from proliferating.

Eye-antennal imaginal discs from different developmental stages carrying ph⁵⁰⁵-clones (A-C).

Differential Expression analysis – comparison of ph⁵⁰⁵-tumor cells (GFP+) with wildtype EA discs.

Generation of ph⁵⁰⁵ mutant clones in wing imaginal discs.

JAK/STAT and JNK (MMP1) pathway activation in ph⁵⁰⁵ clones in wing imaginal discs.