Journal: bioRxiv

Article Title: Glutamine addiction is a therapeutic target to block emergency myelopoiesis

doi: 10.64898/2026.03.26.714544

Figure Lengend Snippet: (A-B) Targeting hematopoietic-specific glutaminolysis during breast cancer development in Ctrl and Gls Δ/Δ mice: (A) breast cancer model with orthotopic implantation of MMTV-PyMT cell line (Py230, 1×10 6 cells); and (B) breast tumor growth over time measured by external palpation (n=16-18; 4 independent experiments; left), with tumor volume at 8 weeks (8-wks) post-orthotopic implantation (n=8-9; 2 independent experiments; right). (C-H) Characterization of non-implanted (NI) and tumor-bearing (PyMT) Ctrl and Gls Δ/Δ mice at 8-wks post-orthotopic implantation: (C) quantification of BM neutrophils (Neu) and granulocyte progenitors (GP) (n=8-9; 2 independent experiments); (D) immunofluorescence imaging of BM GMP patches (stars) and GMP clusters (dotted line) (representative images from 2 independent experiments); (E) transcriptional regulation of HSPCs analyzed by scRNA-seq, with contour density plots of BM LK cells showing tumor-induced loss of granulopoiesis in tumor-bearing Gls Δ/Δ mice; (F) OXPHOS measurement by extracellular flux analysis of LSK, with OCR levels (left) and detailed maximal respiration levels (right) (n=6-9; 6 independent experiments); (G) circulating WBC and Neu plotted as a function of PyMT tumor volume (n=16-18; 3 independent experiments), with simple linear regression reporting slope (S) and goodness of fit (R2); and (H) quantification of intratumoral pro-tumorigenic neutrophil subsets (n=8; 3 independent experiments). O, Oligomycin A; F, FCCP; R/A, Rotenone/Antimycin A; T1, DcTrailR1 - /CD101 - ; T2, DcTrailR1 - /CD101 + ; T3, DcTrailR1 + /CD101 +/- . (I) Targeting myeloid-specific glutaminolysis during breast cancer development in Ctrl and Gls mΔ/Δ mice (n=6-11; 3 independent experiments) with tumor growth over time measured by external palpation (left), and tumor volume at 8 weeks (8-wks) post-orthotopic implantation (right). Data are means ± S.E.M. (B, C, F, H, I) or linear regression with 95% C.I. (G); dots represent individual measurements (G) and circles individual mice (B, C, F, H, I); P. values were obtained by an unpaired t-test (B, C, F, H, I) or a two-tailed test (G). See also Figures S1, S8, S9, and S10.

Article Snippet: MMTV-PyMT mice (FVB/N-Tg(MMTV-PyVT)634Mul/J), Myc-eGfp mice (Myctm1.1Dlev/J), and MRP8-Cre-ires-Gfp mice (B6.Cg-Tg(S100A8-cre,-EGFP)1Ilw/J) were purchased from the Jackson Laboratory.

Techniques: Immunofluorescence, Imaging, Two Tailed Test

Journal: bioRxiv

Article Title: Glutamine addiction is a therapeutic target to block emergency myelopoiesis

doi: 10.64898/2026.03.26.714544

Figure Lengend Snippet: (A-B) Regenerative response in mice treated with the glutamine metabolism inhibitor 6-Diazo-5-oxo-L-norleucine (DON) or PBS vehicle (Veh) control: (A) treatment scheme with daily DON injections of days 6 to 9 post-5FU treatment; and (B) quantification of peripheral blood neutrophils and RBCs (n=10; 3 independent experiments). (C-G) Impact of pharmacological inhibition of glutaminolysis on breast cancer development in Veh and DON-treated WT mice (n=9; 1 independent experiments): (C) breast cancer model with daily DON injections 5 weeks after orthotopic implantation of MMTV-PyMT cell line (Py230, 1×106 cells); (D) breast tumor growth over time measured by external palpation (left), with tumor volume after 14 days of DON treatment; and quantification of (E) BM, (F) peripheral blood and (G) intratumoral myeloid populations after 14 days of DON treatment. (H) Model of glutaminase targeting to impair glutamine-driven OXPHOS and redox balance in myeloid progenitors as a novel therapeutic strategy to suppress tumor-associated myelopoiesis and neutrophil overproduction in breast cancer. Data are means ± S.E.M.; circles represent individual mice; P. values were obtained by an unpaired t-test. See also Figure S1, and S11.

Article Snippet: MMTV-PyMT mice (FVB/N-Tg(MMTV-PyVT)634Mul/J), Myc-eGfp mice (Myctm1.1Dlev/J), and MRP8-Cre-ires-Gfp mice (B6.Cg-Tg(S100A8-cre,-EGFP)1Ilw/J) were purchased from the Jackson Laboratory.

Techniques: Control, Inhibition

Journal: bioRxiv

Article Title: Glutamine addiction is a therapeutic target to block emergency myelopoiesis

doi: 10.64898/2026.03.26.714544

Figure Lengend Snippet: (A-B) Targeting hematopoietic-specific glutaminolysis during breast cancer development in Ctrl and Gls Δ/Δ mice: (A) breast cancer model with orthotopic implantation of MMTV-PyMT cell line (Py230, 1×10 6 cells); and (B) breast tumor growth over time measured by external palpation (n=16-18; 4 independent experiments; left), with tumor volume at 8 weeks (8-wks) post-orthotopic implantation (n=8-9; 2 independent experiments; right). (C-H) Characterization of non-implanted (NI) and tumor-bearing (PyMT) Ctrl and Gls Δ/Δ mice at 8-wks post-orthotopic implantation: (C) quantification of BM neutrophils (Neu) and granulocyte progenitors (GP) (n=8-9; 2 independent experiments); (D) immunofluorescence imaging of BM GMP patches (stars) and GMP clusters (dotted line) (representative images from 2 independent experiments); (E) transcriptional regulation of HSPCs analyzed by scRNA-seq, with contour density plots of BM LK cells showing tumor-induced loss of granulopoiesis in tumor-bearing Gls Δ/Δ mice; (F) OXPHOS measurement by extracellular flux analysis of LSK, with OCR levels (left) and detailed maximal respiration levels (right) (n=6-9; 6 independent experiments); (G) circulating WBC and Neu plotted as a function of PyMT tumor volume (n=16-18; 3 independent experiments), with simple linear regression reporting slope (S) and goodness of fit (R2); and (H) quantification of intratumoral pro-tumorigenic neutrophil subsets (n=8; 3 independent experiments). O, Oligomycin A; F, FCCP; R/A, Rotenone/Antimycin A; T1, DcTrailR1 - /CD101 - ; T2, DcTrailR1 - /CD101 + ; T3, DcTrailR1 + /CD101 +/- . (I) Targeting myeloid-specific glutaminolysis during breast cancer development in Ctrl and Gls mΔ/Δ mice (n=6-11; 3 independent experiments) with tumor growth over time measured by external palpation (left), and tumor volume at 8 weeks (8-wks) post-orthotopic implantation (right). Data are means ± S.E.M. (B, C, F, H, I) or linear regression with 95% C.I. (G); dots represent individual measurements (G) and circles individual mice (B, C, F, H, I); P. values were obtained by an unpaired t-test (B, C, F, H, I) or a two-tailed test (G). See also Figures S1, S8, S9, and S10.

Article Snippet: The syngeneic MMTV-PyMT breast cancer cell line Py230 (ATCC CRL-3279) was used for orthotopic tumor implantation experiments.

Techniques: Immunofluorescence, Imaging, Two Tailed Test

Journal: bioRxiv

Article Title: Glutamine addiction is a therapeutic target to block emergency myelopoiesis

doi: 10.64898/2026.03.26.714544

Figure Lengend Snippet: (A-B) Regenerative response in mice treated with the glutamine metabolism inhibitor 6-Diazo-5-oxo-L-norleucine (DON) or PBS vehicle (Veh) control: (A) treatment scheme with daily DON injections of days 6 to 9 post-5FU treatment; and (B) quantification of peripheral blood neutrophils and RBCs (n=10; 3 independent experiments). (C-G) Impact of pharmacological inhibition of glutaminolysis on breast cancer development in Veh and DON-treated WT mice (n=9; 1 independent experiments): (C) breast cancer model with daily DON injections 5 weeks after orthotopic implantation of MMTV-PyMT cell line (Py230, 1×106 cells); (D) breast tumor growth over time measured by external palpation (left), with tumor volume after 14 days of DON treatment; and quantification of (E) BM, (F) peripheral blood and (G) intratumoral myeloid populations after 14 days of DON treatment. (H) Model of glutaminase targeting to impair glutamine-driven OXPHOS and redox balance in myeloid progenitors as a novel therapeutic strategy to suppress tumor-associated myelopoiesis and neutrophil overproduction in breast cancer. Data are means ± S.E.M.; circles represent individual mice; P. values were obtained by an unpaired t-test. See also Figure S1, and S11.

Article Snippet: The syngeneic MMTV-PyMT breast cancer cell line Py230 (ATCC CRL-3279) was used for orthotopic tumor implantation experiments.

Techniques: Control, Inhibition