Cholesterol efflux protein, ABCA1, supports anticancer functions of myeloid immune cells

Classified as probably not a substantial claim.

Efferocytosis is seen as a major hurdle to using antigen presenting cells in cell-based therapies.

“It is seen as a major hurdle to using antigen presenting cells in cell-based therapies (61–64).”

• cites [61] Ma Y, Jiang T, Zhu X, Xu Y, Wan K, Zhang T, and Xie M. Efferocytosis in dendritic cells: an overlooked immunoregulatory process. Front Immunol. 2024; 15: 1415573. 38835772 PMC11148234 sourceDisputed

  The section on DC maturation and function describes using apoptotic cells to activate antitumor T-cell responses as a promising cancer-vaccine approach. The section on antigen cross-presentation likewise presents DC efferocytosis as informing new immunotherapies and vaccines, rather than as a major hurdle to APC-based therapies. Page numbers are unavailable in our copy. (LLM, conf=0.78)

  Checked by a reasoning model — may be wrong.

  Experimental — accuracy not yet measured
• cites [62] Vaught DB, Stanford JC, and Cook RS. Efferocytosis creates a tumor microenvironment supportive of tumor survival and metastasis. Cancer Cell Microenviron. 2015; 2(1). PMC5734109 29264372 sourcePartly supported

  The Introduction (page not provided) says efferocytosis causes antigen-presenting macrophages to promote immune tolerance and suppress adaptive immune responses. It does not describe efferocytosis as a major hurdle to using antigen-presenting cells in cell-based therapies; that application and degree of difficulty are unsupported. (LLM, conf=0.78)

  Experimental — accuracy not yet measured
• cites [63] Zhou Y, Yao Y, Deng Y, and Shao A. Regulation of efferocytosis as a novel cancer therapy. Cell Commun Signal. 2020; 18(1): 71. 32370748 PMC7199874 sourcePartly supported

  The Immunomodulation section (page unavailable) says efferocytosis suppresses antitumor immune responses, and the Blockade of “eat-me” signaling section links its inhibition to enhanced dendritic-cell cross-presentation. The paper does not describe efferocytosis as a major hurdle to using antigen-presenting cells in cell-based therapies; its discussion of impaired APC function concerns a potential side effect of PS-targeting therapy. (LLM, conf=0.78)

  Experimental — accuracy not yet measured
• cites [64] Mehrotra P, and Ravichandran KS. Drugging the efferocytosis process: concepts and opportunities. Nat Rev Drug Discov. 2022; 21(8): 601– 20. 35650427 PMC9157040 sourceCould not verify

  The Cancer section describes efferocytosis as a challenge for antitumour drug development, and the Infectious diseases section reports reduced antigen presentation by dendritic cells in a specific infection. Neither passage addresses whether efferocytosis is a major hurdle to using antigen-presenting cells in cell-based therapies. Searches for antigen-presenting cells and cell-based therapies found no passage making that connection. (LLM, conf=0.72)

  Experimental — accuracy not yet measured

May combine two claims.

27-hydroxycholesterol works through liver x receptors in myeloid immune cells to suppress T cell proliferation and function, stimulate secretion of pro-tumor extracellular vesicles, and promote tumor and metastatic growth.

“An oxysterol metabolite of cholesterol, 27-hydroxycholesterol, was found to work through the liver x receptors in myeloid immune cells to robustly suppress T cell proliferation and function, and stimulate the secretion of pro-tumor extracellular vesicles, ultimately promoting the growth of tumors and metastatic lesions (17, 20–23).”

• cites [17] Baek AE, Yu Y-RA, He S, Wardell SE, Chang C-Y, Kwon S, The cholesterol metabolite 27 hydroxycholesterol facilitates breast cancer metastasis through its actions on immune cells. Nature Communications. 2017; 8(1): 864. PMC5636879 29021522 sourcePartly supported

  The Results and Discussion report that 27-hydroxycholesterol promotes metastasis through myeloid cells, particularly neutrophils, and is associated with reduced cytotoxic CD8+ T-cell activity. The Discussion links the neutrophil-mediated effect to CXCR2; although it describes 27-hydroxycholesterol as an LXR agonist, it does not establish LXR signaling in myeloid cells as this mechanism. Our copy also does not support the claimed secretion of pro-tumor extracellular vesicles. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [20] Baek AE, Krawczynska N, Das Gupta A, Dvoretskiy SV, You S, Park J, The cholesterol metabolite 27-hydroxycholesterol increases the secretion of extracellular vesicles which promote breast cancer progression. Endocrinology. 2021. PMC8197285 33959755 sourcePartly supported

  The Abstract reports that 27HC acts through LXRs in myeloid cells to suppress anticancer T-cell activity, while the Results and Discussion report increased EV secretion and tumor growth and metastasis from 27HC-induced EVs. The Discussion says the mechanism by which 27HC stimulates EV secretion remains to be determined, so the paper does not establish that LXR signaling mediates that secretion. Page numbers are not available in our copy. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [21] Das Gupta A, Park J, Sorrells JE, Kim H, Krawczynska N, Pradeep D, 27-Hydroxycholesterol Enhances Secretion of Extracellular Vesicles by ROS-Induced Dysregulation of Lysosomes. Endocrinology. 2024. PMC11448339 39298675 sourcePartly supported

  The introductory passage reports that 27HC acts through liver X receptors in myeloid cells to suppress T-cell expansion, and the Discussion reports that 27HC-induced extracellular vesicles promote tumor growth and metastasis. But the paper attributes increased vesicle secretion to mitochondrial dysfunction, oxidative stress, and altered lysosomal function; it does not establish liver X receptors as the mechanism for that secretion or the resulting tumor growth. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [22] Ma L, Wang L, Nelson AT, Han C, He S, Henn MA, 27-Hydroxycholesterol acts on myeloid immune cells to induce T cell dysfunction, promoting breast cancer progression. Cancer letters. 2020; 493: 266– 83. 32861706 PMC7572761 sourcePartly supported

  The Results report that 27-hydroxycholesterol acts through LXRs in myeloid cells to impair T-cell expansion, and that it increases metastatic burden in mouse models. But the Results section on macrophage secretion found no difference in the effects of extracellular vesicles from treated and untreated macrophages; it concludes that vesicles are unlikely to mediate the observed T-cell suppression. The paper therefore does not support the attributed claim that 27-hydroxycholesterol stimulates secretion of pro-tumor extracellular vesicles. (LLM, conf=0.90)

  Experimental — accuracy not yet measured
• cites [23] Ma L, Vidana Gamage HE, Tiwari S, Han C, Henn MA, Krawczynska N, The Liver X Receptor Is Selectively Modulated to Differentially Alter Female Mammary Metastasis-associated Myeloid Cells. Endocrinology. 2022; 163(7). PMC9188661 35569056 sourcePartly supported

  The Abstract says 27HC’s effects on myeloid cells require LXR and impair T-cell expansion and function; it also describes protumoral and prometastatic effects. The cited paper does not report that 27HC stimulates secretion of pro-tumor extracellular vesicles (searched: extracellular vesicles, vesicles, exosomes). (LLM, conf=0.88)

  Experimental — accuracy not yet measured

May combine two claims.

Myeloid cells are required for antigen presentation, T cell support and effective anti-tumor responses.

“However, myeloid cells are required for antigen presentation, T cell support and effective anti-tumor responses (15).”

• cites [15] Mantovani A, Allavena P, Marchesi F, and Garlanda C. Macrophages as tools and targets in cancer therapy. Nat Rev Drug Discov. 2022. PMC9380983 35974096 sourcePartly supported

  The CD40 section (page unavailable) identifies macrophages as antigen-presenting cells that can stimulate T cell-mediated immunity, and the Introduction describes their potential to activate antitumour lymphoid responses. These passages support a beneficial role for some myeloid cells, but do not establish that myeloid cells are required for antigen presentation, T cell support, or effective antitumour responses generally. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

May combine two claims.

Statins have high first-pass metabolism, allowing tumors to escape and produce their own cholesterol.

“However, statins have a high first-pass metabolism, allowing for tumors to escape and produce their own cholesterol (11, 12).”

• cites [11] Bjarnadottir O, Romero Q, Bendahl PO, Jirstrom K, Ryden L, Loman N, Targeting HMG-CoA reductase with statins in a window-of-opportunity breast cancer trial. Breast cancer research and treatment. 2013; 138(2): 499– 508. 23471651 sourceNot checked — no full text of the source
• cites [12] Kimbung S, Lettiero B, Feldt M, Bosch A, and Borgquist S. High expression of cholesterol biosynthesis genes is associated with resistance to statin treatment and inferior survival in breast cancer. Oncotarget. 2016; 7(37): 59640– 51. 27458152 PMC5312337 sourcePartly supported

  The Discussion (page unavailable) says most orally administered statins are unlikely to reach the concentrations used in vitro outside the liver. It also reports that less-sensitive breast cancer cells strongly induced cholesterol-biosynthesis genes after statin treatment. The paper does not establish high first-pass metabolism or show that it allows tumors to escape treatment by producing their own cholesterol. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

May combine two claims.

In TNBC patients, statin use is associated with improved overall survival and breast cancer specific survival.

“This association is also true for TNBC patients taking statins, demonstrating improved overall survival and breast cancer specific survival (10).”

• cites [10] Nowakowska MK, Lei X, Thompson MT, Shaitelman SF, Wehner MR, Woodward WA, Association of statin use with clinical outcomes in patients with triple-negative breast cancer. Cancer. 2021; 127(22): 4142– 50. 34342892 PMC11912801 sourceSupported

May combine two claims.

NR0B2 suppresses several aspects of the inflammasome in myeloid cells, resulting in decreased expansion of regulatory T cells.

“The negative regulator of cholesterol catabolism, NR0B2, was found to suppress several aspects of the inflammasome in myeloid cells, resulting in decreased expansion of regulatory T cells (Tregs) (24, 25).”

• cites [24] Shahoei SH, Kim YC, Cler SJ, Ma L, Anakk S, Kemper JK, and Nelson ER. Small Heterodimer Partner Regulates Dichotomous T Cell Expansion by Macrophages. Endocrinology. 2019; 160(7): 1573– 89. 31050726 PMC6549582 sourceNot checked — no full text of the source
• cites [25] Vidana Gamage HE, Shahoei SH, Wang Y, Jacquin E, Weisser E, Bautista RO, NR0B2 re-educates myeloid immune cells to reduce regulatory T cell expansion and progression of breast and other solid tumors. Cancer letters. 2024; 597: 217042. 38908543 PMC11892351 sourceSupported

May combine two claims.

These trends are likely driven by ABCA1 expression extrinsic to T cells (i.e., in MCs), as cholesterol efflux has been shown to impair T cell expansion and activity.

“These trends are likely driven by ABCA1 expression extrinsic to T cells (ie: in MCs), as cholesterol efflux has been shown to impair T cell expansion and activity (39, 40).”

• cites [39] Bensinger SJ, Bradley MN, Joseph SB, Zelcer N, Janssen EM, Hausner MA, LXR signaling couples sterol metabolism to proliferation in the acquired immune response. Cell. 2008; 134(1): 97– 111. 18614014 PMC2626438 sourcePartly supported

  The Results section reports that LXR activation reduces T-cell proliferation through an ABCG1-dependent change in sterol metabolism; it also links LXR signaling to antigen-driven T-cell responses. But the Discussion describes sterol efflux or redistribution as a likely mechanism, not cholesterol efflux as an established cause. The paper does not show that ABCA1 expression outside T cells drives the citing authors’ trends; its experiments instead find that loss of ABCA1 does not alter the T-cell response to LXR agonists. (LLM, conf=0.85)

  Experimental — accuracy not yet measured
• cites [40] Joseph SB, Castrillo A, Laffitte BA, Mangelsdorf DJ, and Tontonoz P. Reciprocal regulation of inflammation and lipid metabolism by liver X receptors. Nat Med. 2003; 9(2): 213– 9. 12524534 sourceNot checked — no full text of the source

May combine two claims.

There was enrichment in the PI3K-Akt signaling pathway as well as the Rap1 signaling pathway which has crosstalk with the PI3K-Akt pathway.

“We observed enrichment in the PI3K-Akt signaling pathway as well as the Rap1 signaling pathway which has crosstalk with the PI3K-Akt pathway (Fig. 7B) (41).”

• cites [41] Zhang YL, Wang RC, Cheng K, Ring BZ, and Su L. Roles of Rap1 signaling in tumor cell migration and invasion. Cancer Biol Med. 2017; 14(1): 90– 9. 28443208 PMC5365179 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

ABCA1 is a protein involved in cholesterol efflux and translocation from the inner leaflet to the outer leaflet of the plasma membrane.

“we identified ABCA1, a protein involved in cholesterol efflux and translocation from the inner leaflet to the outer leaflet of the plasma membrane (4, 27).”

• cites [4] Wang Y, Bendre SV, Krauklis SA, Steelman AJ, and Nelson ER. Role of protein regulators of cholesterol homeostasis in immune modulation and cancer pathophysiology. Endocrinology. 2025; Accepted. PMC11878532 39951497 sourcePartly supported

  The passage on ABCA1 and reverse cholesterol transport says ABCA1 transports cellular cholesterol to apolipoproteins at the cell surface, supporting its role in cholesterol efflux. Another passage says it shuttles cholesterol from the cytoplasm to the cell surface. Neither passage specifies translocation from the inner to the outer leaflet of the plasma membrane. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [27] Lawn RM, Wade DP, Garvin MR, Wang X, Schwartz K, Porter JG, The Tangier disease gene product ABC1 controls the cellular apolipoprotein-mediated lipid removal pathway. J Clin Invest. 1999; 104(8): R25– 31. 10525055 PMC481052 sourcePartly supported

  The Discussion (p. R25) identifies ABC1 as pivotal to apolipoprotein-mediated cholesterol efflux and says its cell-surface expression is consistent with lipid transport across the plasma membrane. It does not specify translocation of cholesterol from the inner to the outer leaflet; that mechanism is not established by the cited passages. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

HMVECs cocultured with BMDMs from myeloid cell conditional ABCA1 knockout mice had increased proliferation.

“Similar to our findings, HMVECs cocultured with BMDMs from myeloid cell conditional ABCA1 knockout mice had increased proliferation (59).”

• cites [59] Sene A, Khan AA, Cox D, Nakamura RE, Santeford A, Kim BM, Impaired cholesterol efflux in senescent macrophages promotes age-related macular degeneration. Cell Metab. 2013; 17(4): 549– 61. 23562078 PMC3640261 sourcePartly supported

  The Results section reports that HMVECs cocultured with macrophages from myeloid-conditional Abca1 knockout mice showed greater proliferation than those cocultured with wild-type macrophages. However, the Vascular Endothelial Cell Proliferation Assay specifies peritoneal macrophages, not bone-marrow-derived macrophages (BMDMs). (LLM, conf=0.90)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

Previous work found that double knockout of ABCA1 and ABCG1 in macrophages resulted in decreased migratory capacity when stimulated with C5a, ascribed to Rac1 activation.

“Previous work found that double knockout of ABCA1 and ABCG1 in macrophages resulted in decreased migratory capacity when stimulated with C5a (compliment component 5a), compared to wildtype macrophages, ascribed to Rac1 activation (55).”

• cites [55] Pagler TA, Wang M, Mondal M, Murphy AJ, Westerterp M, Moore KJ, Deletion of ABCA1 and ABCG1 impairs macrophage migration because of increased Rac1 signaling. Circ Res. 2011; 108(2): 194– 200. 21148432 PMC3097897 sourceSupported

May combine two claims.

Efferocytosis does not culminate in antigen presentation and instead evokes a highly immune suppressive macrophage phenotype.

“Efferocytosis does not culminate in antigen presentation and instead evokes a highly immune suppressive macrophage phenotype (35, 36).”

• cites [35] Moon B, Yang S, Moon H, Lee J, and Park D. After cell death: the molecular machinery of efferocytosis. Exp Mol Med. 2023; 55(8): 1644– 51. 37612408 PMC10474042 sourcePartly supported

  The Introduction (page unavailable) says engulfment of apoptotic cells generally induces anti-inflammatory responses or is immunologically silent. The Degradation of apoptotic cells section (page unavailable) reports increased production of anti-inflammatory mediators, including IL-10. These passages support an anti-inflammatory response, but do not establish that efferocytosis never culminates in antigen presentation or always produces a highly immunosuppressive macrophage phenotype. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [36] Elliott MR, Koster KM, and Murphy PS. Efferocytosis Signaling in the Regulation of Macrophage Inflammatory Responses. J Immunol. 2017; 198(4): 1387– 94. 28167649 PMC5301545 sourcePartly supported

  The Introduction’s “Cell death in inflamed tissues” section says apoptotic cells can suppress macrophage inflammatory responses and that efferocytosis promotes resolution, supporting the immunosuppressive aspect. The cited paper does not establish the separate, categorical claim that efferocytosis does not culminate in antigen presentation; searches for antigen presentation and related terms found no supporting passage in our copy. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Previous work demonstrated that LXR induction of ABCA1 reduces angiogenesis, presumably by decreasing lipid raft mediated stabilization of the VEGF-VEGFR complex, within vascular endothelial cells.

“Previous work has demonstrated that LXR induction of ABCA1 reduces angiogenesis, presumably by decreasing lipid raft mediated stabilization of the VEGF-VEGFR complex, within vascular endothelial cells (56).”

• cites [56] Noghero A, Perino A, Seano G, Saglio E, Lo Sasso G, Veglio F, Liver X receptor activation reduces angiogenesis by impairing lipid raft localization and signaling of vascular endothelial growth factor receptor-2. Arterioscler Thromb Vasc Biol. 2012; 32(9): 2280– 8. 22723445 sourcePartly supported

  The Abstract and Discussion report that LXR activation reduces angiogenesis in endothelial cells and impairs VEGFR2 localization and signaling in lipid rafts. The Discussion also says LXR activation increases both ABCA1 and ABCG1; it does not establish that ABCA1 induction alone causes the anti-angiogenic effect or that lipid rafts stabilize a VEGF–VEGFR complex. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Angiogenesis is increased in zebrafish embryos lacking ABCA1 and ABCG1.

“Similarly, angiogenesis is increased in zebrafish embryos lacking ABCA1 and ABCG1 (57).”

• cites [57] Fang L, Choi SH, Baek JS, Liu C, Almazan F, Ulrich F, Control of angiogenesis by AIBP-mediated cholesterol efflux. Nature. 2013; 498(7452): 118– 22. 23719382 PMC3760669 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Loss of ABCA1 leads to a buildup of cholesterol within the cell, engaging cholesterol-related feedback mechanisms.

“This would be expected since loss of ABCA1 would lead to a buildup of cholesterol within the cell, engaging cholesterol-related feedback mechanisms (45).”

• cites [45] Ma L, Cho W, and Nelson ER. Our evolving understanding of how 27-hydroxycholesterol influences cancer. Biochem Pharmacol. 2022; 196: 114621. 34043965 PMC8611110 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Macrophages preferentially efferocytose cancer cells, and significant efforts are being made to inhibit efferocytosis therapeutically.

“Indeed, macrophage cell therapy has faced a significant hurdle in that macrophages preferentially efferocytose cancer cells, and significant efforts are being made to inhibit efferocytosis therapeutically (37, 38).”

• cites [37] Werfel TA, and Cook RS. Efferocytosis in the tumor microenvironment. Semin Immunopathol. 2018; 40(6): 545– 54. 30187085 PMC6223858 sourcePartly supported

  The “Therapeutic targeting of efferocytosis in the setting of cancer” section describes inhibitors being developed to block efferocytosis, supporting the therapeutic-efforts claim. But the “Mechanisms of efferocytosis” section says phagocytes preferentially identify apoptotic cells rather than healthy cells; it does not say macrophages preferentially efferocytose cancer cells. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [38] Tsai WH, Chang SC, Lin YC, and Hsu HC. CX3CL1(+) Microparticles-Induced MFG-E8 Enhances Apoptotic Cell Clearance by Alveolar Macrophages. Cells. 2021; 10(10). PMC8533702 34685562 sourcePartly supported

  The Results (§3.1 and §3.5; page unavailable) show alveolar macrophages engulfing apoptotic leukemic NB4 cells more readily than viable NB4 cells. They do not establish a preference for cancer cells over other cell types or discuss macrophage cell therapy. The Discussion (§4; page unavailable) focuses on enhancing efferocytosis, not therapeutic efforts to inhibit it. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Altering membrane cholesterol might affect lipid raft formation and subsequent PI3K signaling, or membrane-bound cholesterol can directly activate certain downstream signaling proteins such as Akt or Dvl2 (Wnt signaling).

“Enrichment in the PI3K-Akt and Wnt signaling pathways was of interest given reports that altering membrane cholesterol might affect lipid raft formation and subsequent PI3K signaling (42–44), or that membrane-bound cholesterol can directly activate certain downstream signaling proteins such as Akt or Dvl2 (Wnt signaling).”

• cites [42] Erazo-Oliveras A, Munoz-Vega M, Mlih M, Thiriveedi V, Salinas ML, Rivera-Rodriguez JM, Mutant APC reshapes Wnt signaling plasma membrane nanodomains by altering cholesterol levels via oncogenic beta-catenin. Nat Commun. 2023; 14(1): 4342. 37468468 PMC10356786 sourcePartly supported

  The Discussion reports that membrane free cholesterol interacts with Dvl and promotes its scaffolding function in Wnt signaling; the Results specifically measure Dvl1. The cited copy does not establish the sentence’s PI3K–Akt claim or identify Dvl2 as the protein studied. Its account of Dvl also describes cholesterol-dependent recruitment and interactions, rather than direct activation in isolation. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [43] Sheng R, Kim H, Lee H, Xin Y, Chen Y, Tian W, Cholesterol selectively activates canonical Wnt signalling over non-canonical Wnt signalling. Nat Commun. 2014; 5: 4393. 25024088 PMC4100210 sourcePartly supported

  The Results section ('Dvl PDZ domain specifically binds cholesterol') reports that Dvl2 binds cholesterol-containing membranes, and the Discussion says this binding promotes Dvl2’s role in canonical Wnt signaling. The paper does not establish direct activation of Akt or a lipid-raft-mediated PI3K effect; its Discussion says Dvl’s cholesterol-dependent activity need not involve lipid-raft formation. The supplied copy gives no page numbers for these passages. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [44] Sheng R, Chen Y, Yung Gee H, Stec E, Melowic HR, Blatner NR, Cholesterol modulates cell signaling and protein networking by specifically interacting with PDZ domain-containing scaffold proteins. Nat Commun. 2012; 3: 1249. 23212378 PMC3526836 sourcePartly supported

  The Introduction says cholesterol contributes to lipid-raft formation, and the Discussion describes cholesterol-rich rafts as proposed signaling platforms. Neither section links those rafts to PI3K signaling. The paper demonstrates cholesterol binding to PDZ-domain scaffold proteins, not direct activation of Akt or Dvl2. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

AKT3 has been implicated in atherosclerosis and tissue remodeling.

“Less is known about the effects of AKT3 on MΦ function, although it has been implicated in atherosclerosis (47, 48) and tissue remodeling (49).”

• cites [47] Zhang L, Altemus J, Ding L, Cherepanova O, Byzova TV, and Podrez EA. Enhanced Akt3 kinase activity reduces atherosclerosis in hyperlipidemic mice in a gender-dependent manner. J Biol Chem. 2023; 299(12): 105425. 37926285 PMC10716582 sourceSupported
• cites [48] Ding L, Biswas S, Morton RE, Smith JD, Hay N, Byzova TV, Akt3 deficiency in macrophages promotes foam cell formation and atherosclerosis in mice. Cell Metab. 2012; 15(6): 861– 72. 22632897 PMC3372639 sourceSupported
• cites [49] Gu S, Dai H, Zhao X, Gui C, and Gui J. AKT3 deficiency in M2 macrophages impairs cutaneous wound healing by disrupting tissue remodeling. Aging (Albany NY). 2020; 12(8): 6928– 46. 32291381 PMC7202485 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Angiogenesis is associated with the classical M2 polarization state of macrophages.

“Angiogenesis is associated with the classical M2 polarization state of macrophages (60).”

• cites [60] Corliss BA, Azimi MS, Munson JM, Peirce SM, and Murfee WL. Macrophages: An Inflammatory Link Between Angiogenesis and Lymphangiogenesis. Microcirculation. 2016; 23(2): 95– 121. 26614117 PMC4744134 sourcePartly supported

  The “Macrophage phenotypic diversity” section links M2 macrophages to cytokines that promote angiogenesis, supporting the association. But it identifies M1—not M2—as the classically activated phenotype; M2 is alternatively activated. The supplied passages do not specify page numbers. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Endothelial cell knockout of ABCA1 alone did not result in alterations in new vessel sprouting in a murine model of atherosclerosis, while knockout of ABCG1 did.

“However, endothelial cell knockout of ABCA1 alone did not result in alterations in new vessel sprouting in a murine model of atherosclerosis, while knockout of ABCG1 did (58).”

• cites [58] Westerterp M, Tsuchiya K, Tattersall IW, Fotakis P, Bochem AE, Molusky MM, Deficiency of ATP-Binding Cassette Transporters A1 and G1 in Endothelial Cells Accelerates Atherosclerosis in Mice. Arterioscler Thromb Vasc Biol. 2016; 36(7): 1328– 37. 27199450 PMC4919153 sourcePartly supported

  The Results section ('EC-Abca1 and Abcg1 deficiency enhance new vessel sprouting in aortic rings'; page not provided) reports increased sprouting with endothelial Abcg1 knockout, but not Abca1 knockout. That finding came from an ex vivo aortic-ring assay, not a measurement of sprouting in atherosclerotic lesions; the paper reports no difference in new blood vessels in those lesions. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Cholesterol is required to support the rapid clonal expansion of T cells post-activation.

“Cholesterol is required to support the rapid clonal expansion of T cells post-activation (39, 40); plasma membrane cholesterol abundance being required for T-cell receptor clustering and signaling (69).”

• cites [39] Bensinger SJ, Bradley MN, Joseph SB, Zelcer N, Janssen EM, Hausner MA, LXR signaling couples sterol metabolism to proliferation in the acquired immune response. Cell. 2008; 134(1): 97– 111. 18614014 PMC2626438 sourceSupported
• cites [40] Joseph SB, Castrillo A, Laffitte BA, Mangelsdorf DJ, and Tontonoz P. Reciprocal regulation of inflammation and lipid metabolism by liver X receptors. Nat Med. 2003; 9(2): 213– 9. 12524534 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

No approaches targeting efferocytosis have thus far been clinically approved.

“While some progress has been made with respect to the Fractalkine receptor (CX3C motif chemokine receptor 1), Axl, MerTK, CD47 and Tim4/3, no approaches have thus far been clinically approved (64).”

• cites [64] Mehrotra P, and Ravichandran KS. Drugging the efferocytosis process: concepts and opportunities. Nat Rev Drug Discov. 2022; 21(8): 601– 20. 35650427 PMC9157040 sourcePartly supported

  Table 1 (Approaches to target the eating phase; page unavailable) lists therapeutics targeting Axl, MerTK, CD47 and TIM4/3 as being in clinical trials, supporting the distinction between progress in trials and clinical approval. The Introduction (page unavailable) also says an autologous apoptotic-cell approach is already in use. The review does not establish the broader claim that no approach targeting efferocytosis has been clinically approved. (LLM, conf=0.78)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

T cell specific knockout of ABCA1 and ABCG1 mice had decreased T cell numbers but increased activation, resulting in decreased atherosclerosis and aortic inflammation.

“T cell specific knockout of ABCA1 and ABCG1 mice had decreased T cell numbers but increased activation, resulting in decreased atherosclerosis and aortic inflammation (71).”

• cites [71] Bazioti V, La Rose AM, Maassen S, Bianchi F, de Boer R, Halmos B, T cell cholesterol efflux suppresses apoptosis and senescence and increases atherosclerosis in middle aged mice. Nat Commun. 2022; 13(1): 3799. 35778407 PMC9249754 sourcePartly supported

  The Results sections on peripheral T cells and activation report fewer T cells and increased activation after T cell-specific Abca1/Abcg1 deletion (page unavailable). The section on middle-aged Ldlr−/− mice reports smaller atherosclerotic lesions and changes consistent with reduced aortic inflammation, but attributes these effects primarily to increased T cell apoptosis and reduced inflammatory signaling—not increased activation. The paper also reports no change in lesion size in the other Ldlr−/− mice studied, so the claim omits an important age qualification. (LLM, conf=0.90)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

A previous report indicated that myeloid cell loss of ABCA1 decreased B16-F10 melanoma and MB49 bladder tumor growth.

“However, a previous report indicated that myeloid cell loss of ABCA1 decreased B16-F10 melanoma and MB49 bladder tumor growth (75).”

• cites [75] Zamanian-Daryoush M, Lindner DJ, DiDonato JA, Wagner M, Buffa J, Rayman P, Myeloid-specific genetic ablation of ATP-binding cassette transporter ABCA1 is protective against cancer. Oncotarget. 2017; 8(42): 71965– 80. 29069761 PMC5641104 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Plasma membrane cholesterol abundance is required for T-cell receptor clustering and signaling.

“Cholesterol is required to support the rapid clonal expansion of T cells post-activation (39, 40); plasma membrane cholesterol abundance being required for T-cell receptor clustering and signaling (69).”

• cites [69] Yang W, Bai Y, Xiong Y, Zhang J, Chen S, Zheng X, Potentiating the antitumour response of CD8(+) T cells by modulating cholesterol metabolism. Nature. 2016; 531(7596): 651– 5. 26982734 PMC4851431 sourceSupported

May combine two claims.

Apoptotic cells increase ABCA1, which is required for the secretion of find me signals.

“Interestingly, apoptotic cells increase ABCA1, which is required for the secretion of ‘find me’ signals (65, 66).”

• cites [65] Fond AM, Lee CS, Schulman IG, Kiss RS, and Ravichandran KS. Apoptotic cells trigger a membrane-initiated pathway to increase ABCA1. J Clin Invest. 2015; 125(7): 2748– 58. 26075824 PMC4563683 sourcePartly supported

  The Results report that apoptotic cells increase ABCA1 expression in primary macrophages. The cited paper does not establish that ABCA1 is required for secretion of ‘find me’ signals; it discusses ABCA1 in relation to cholesterol efflux instead (Results and Discussion; page numbers unavailable). (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [66] Chen W, Li L, Wang J, Zhang R, Zhang T, Wu Y, The ABCA1-efferocytosis axis: A new strategy to protect against atherosclerosis. Clin Chim Acta. 2021; 518: 1– 8. 33741356 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Colorectal cancer APC oncogene increased plasma membrane cholesterol and thus WNT signaling hubs (lipid rafts).

“Another example comes from colorectal cancer where the APC oncogene increased plasma membrane cholesterol and thus WNT signaling hubs (lipid rafts) (38).”

• cites [38] Tsai WH, Chang SC, Lin YC, and Hsu HC. CX3CL1(+) Microparticles-Induced MFG-E8 Enhances Apoptotic Cell Clearance by Alveolar Macrophages. Cells. 2021; 10(10). PMC8533702 34685562 sourceCould not verify

  The Introduction and Results section 3.2 discuss acute promyelocytic leukemia and macrophage clearance of apoptotic cells, not colorectal cancer. Searches for APC, plasma-membrane cholesterol, WNT signaling, and lipid rafts found no passage addressing the claim in our copy; page numbers were unavailable. (LLM, conf=0.76)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S1 (asserted), S3 (load-bearing). May combine two claims.

Ovarian tumor cells secrete some factor that upregulates ABCA1 and ABCG1 in macrophages, which then polarized them into a tumor-associated or M2-like state.

“Another report suggested that ovarian tumor cells secrete some factor that upregulates ABCA1 and ABCG1 in macrophages, which then polarized them into a tumor-associated or “M2-like” state (77).”

• cites [77] Goossens P, Rodriguez-Vita J, Etzerodt A, Masse M, Rastoin O, Gouirand V, Membrane Cholesterol Efflux Drives Tumor-Associated Macrophage Reprogramming and Tumor Progression. Cell Metab. 2019; 29(6): 1376– 89 e4. 30930171 sourcePartly supported

  The Discussion says ovarian cancer cells promote macrophage cholesterol efflux and TAM reprogramming, and suggests tumor-produced hyaluronan as a possible factor. But the Limitations section reports no increase in Abca1 or Abcg1 mRNA after exposure to tumor-cell conditioned medium; the mechanism increasing transporter activity remains unresolved. The claim therefore overstates the evidence by saying the factor upregulates ABCA1 and ABCG1. (LLM, conf=0.90)

  Experimental — accuracy not yet measured

May combine two claims.

Tumors grown in hypercholesterolemic mice (ApoE−/−) have increased PI3K and AKT signaling, and treatment with a PI3K inhibitor attenuated growth.

“One example of this comes from tumors grown in hypercholesterolemic mice (ApoE−/−), which have increased PI3K and AKT signaling, and treatment with a PI3K inhibitor was able to attenuate growth (78).”

• cites [78] Alikhani N, Ferguson RD, Novosyadlyy R, Gallagher EJ, Scheinman EJ, Yakar S, and LeRoith D. Mammary tumor growth and pulmonary metastasis are enhanced in a hyperlipidemic mouse model. Oncogene. 2013; 32(8): 961– 7. 22469977 PMC4063440 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Previous reports indicated that PI3K modulates efferocytosis.

“Indeed, previous reports have indicated that PI3K modulates efferocytosis (80)”

• cites [80] Tao H, Yancey PG, Babaev VR, Blakemore JL, Zhang Y, Ding L, Macrophage SR-BI mediates efferocytosis via Src/PI3K/Rac1 signaling and reduces atherosclerotic lesion necrosis. J Lipid Res. 2015; 56(8): 1449– 60. 26059978 PMC4513986 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Within macrophages, ABCA1 seems to protect from the cellular stress associated with engulfing apoptotic bodies, although ABCG1 appeared to be more predominant in this role.

“Within macrophages, ABCA1 seems to protect from the cellular stress associated with engulfing apoptotic bodies, although ABCG1 appeared to be more predominant in this role (67).”

• cites [67] Yvan-Charvet L, Pagler TA, Seimon TA, Thorp E, Welch CL, Witztum JL, ABCA1 and ABCG1 protect against oxidative stress-induced macrophage apoptosis during efferocytosis. Circ Res. 2010; 106(12): 1861– 9. 20431058 PMC2995809 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Cancer cells are known to stimulate efferocytotic pathways.

“The effects were more pronounced when using apoptotic neutrophils as bait compared to cancer cells, perhaps because cancer cells are known to stimulate efferocytotic pathways (68).”

• cites [68] Tajbakhsh A, Gheibi Hayat SM, Movahedpour A, Savardashtaki A, Loveless R, Barreto GE, The complex roles of efferocytosis in cancer development, metastasis, and treatment. Biomed Pharmacother. 2021; 140: 111776. 34062411 sourcePartly supported

  The Abstract describes efferocytosis of apoptotic tumor cells, and section 2.3.5 discusses tumor-cell molecules involved in their engulfment. This supports the claim for apoptotic cancer cells, but the cited paper does not establish that cancer cells generally stimulate efferocytotic pathways; section 2 also notes that tumor-cell CD47 can inhibit recognition by immune cells. No page numbers were supplied for these passages. (LLM, conf=0.82)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Cholesterol homeostasis has been implicated in tissue resident memory CD8 T cells.

“Cholesterol homeostasis has also been implicated in tissue resident memory CD8 T cells (70).”

• cites [70] Reina-Campos M, Heeg M, Kennewick K, Mathews IT, Galletti G, Luna V, Metabolic programs of T cell tissue residency empower tumour immunity. Nature. 2023; 621(7977): 179– 87. 37648857 PMC11238873 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Certain PZD-domain containing proteins such as Dvl2 can be directly activated by cholesterol within the inner leaflet of the plasma membrane.

“Recent reports suggest that certain PZD-domain containing proteins such as Dvl2 can be directly activated by cholesterol within the inner leaflet of the plasma membrane (43, 44, 82–84).”

• cites [43] Sheng R, Kim H, Lee H, Xin Y, Chen Y, Tian W, Cholesterol selectively activates canonical Wnt signalling over non-canonical Wnt signalling. Nat Commun. 2014; 5: 4393. 25024088 PMC4100210 sourceSupported
• cites [44] Sheng R, Chen Y, Yung Gee H, Stec E, Melowic HR, Blatner NR, Cholesterol modulates cell signaling and protein networking by specifically interacting with PDZ domain-containing scaffold proteins. Nat Commun. 2012; 3: 1249. 23212378 PMC3526836 sourcePartly supported

  The Results (‘Many PDZ domains bind cholesterol’; ‘NHERF1-cholesterol binding is essential for its activity’; page not provided) show that several PDZ domains bind cholesterol in plasma-membrane-mimetic vesicles and that cholesterol binding is important for NHERF1-mediated regulation of CFTR. The source does not identify Dvl2 as one of those proteins or show that cholesterol directly activates Dvl2. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [82] Sheng R, Jung DJ, Silkov A, Kim H, Singaram I, Wang ZG, Lipids Regulate Lck Protein Activity through Their Interactions with the Lck Src Homology 2 Domain. J Biol Chem. 2016; 291(34): 17639– 50. 27334919 PMC5016160 sourcePartly supported

  The Abstract (page unavailable) reports that PDZ domains bind plasma-membrane lipids and that this binding is important for their host proteins’ function. The References list studies of cholesterol interactions with PDZ proteins and cholesterol-dependent Wnt signaling, but this paper does not state that cholesterol directly activates Dvl2 or locate that interaction in the inner leaflet. (LLM, conf=0.78)

  Experimental — accuracy not yet measured
• cites [83] Liu SL, Sheng R, Jung JH, Wang L, Stec E, O’Connor MJ, Orthogonal lipid sensors identify transbilayer asymmetry of plasma membrane cholesterol. Nat Chem Biol. 2017; 13(3): 268– 74. 28024150 PMC5912897 sourcePartly supported

  The Results section ('Stimulus-dependent redistribution of PM cholesterol'; page unavailable) reports that an increase in inner-leaflet cholesterol correlates with Dvl2 recruitment to the plasma membrane, and cites an earlier report of cholesterol-dependent Dvl activation. It does not show that inner-leaflet cholesterol directly activates Dvl2 or establish the proposed PDZ-domain mechanism. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites [84] Liu SL, Wang ZG, Hu Y, Xin Y, Singaram I, Gorai S, Quantitative Lipid Imaging Reveals a New Signaling Function of Phosphatidylinositol-3,4-Bisphophate: Isoform- and Site-Specific Activation of Akt. Molecular cell. 2018; 71(6): 1092– 104 e5. 30174291 PMC6214670 sourceCould not verify

  The Introduction and Discussion address phosphoinositide signaling and Akt activation, not cholesterol-dependent activation of Dvl2. Searches for Dvl2, PDZ proteins, cholesterol, and the inner leaflet found only a related paper title in the References; our copy does not state the claimed mechanism. (LLM, conf=0.76)

  Experimental — accuracy not yet measured

May combine two claims.

Dendritic cells lacking both ABCA1 and ABCG1 have enhanced inflammasome activation, ultimately resulting in enhanced T cell activation and polarization to the Th1 and Th17 types.

“Dendritic cells lacking both ABCA1 and ABCG1 have enhanced inflammasome activation, ultimately resulting in enhanced T cell activation and polarization to the Th1 and Th17 types (74).”

• cites [74] Westerterp M, Gautier EL, Ganda A, Molusky MM, Wang W, Fotakis P, Cholesterol Accumulation in Dendritic Cells Links the Inflammasome to Acquired Immunity. Cell Metab. 2017; 25(6): 1294– 304 e6. 28479366 PMC5514787 sourceSupported

May combine two claims.

Dendritic cells treated with a statin (zoledronic acid) increased ABCA1 which enhanced extracellular release of the phospho-antigen isopentenyl pyrophosphate, ultimately activating Vγ9Vδ2 T cells.

“Using a somewhat contrived system, dendritic cells treated with a statin (zoledronic acid) increased ABCA1 which enhanced extracellular release of the phospho-antigen isopentenyl pyrophosphate, ultimately activating Vγ9Vδ2 T cells (72).”

• cites [72] Castella B, Kopecka J, Sciancalepore P, Mandili G, Foglietta M, Mitro N, The ATP-binding cassette transporter A1 regulates phosphoantigen release and Vgamma9Vdelta2 T cell activation by dendritic cells. Nat Commun. 2017; 8: 15663. 28580927 PMC5465356 sourcePartly supported

  The Results section (“ZA increases IPP efflux via LXRα-mediated Abca1 activation”) and Discussion support the chain from zoledronic acid (ZA) treatment to increased ABCA1, extracellular IPP release, and Vγ9Vδ2 T-cell activation. But the claim incorrectly calls ZA a statin: the paper identifies simvastatin as the statin and reports that it reduces ABCA1 expression and extracellular IPP release. Page numbers are not provided in our copy. (LLM, conf=0.93)

  Experimental — accuracy not yet measured

May combine two claims.

Knockdown of ABCG1 resulted in inhibited tumor growth, but in a different study this required a physiologic challenge (western diet, age or genetic model of hypercholesterolemia).

“Knockdown of the related transporter ABCG1 also resulted in inhibited tumor growth (75), but in a different study this required a physiologic challenge (western diet, age or genetic model of hypercholesterolemia) (76).”

• cites [75] Zamanian-Daryoush M, Lindner DJ, DiDonato JA, Wagner M, Buffa J, Rayman P, Myeloid-specific genetic ablation of ATP-binding cassette transporter ABCA1 is protective against cancer. Oncotarget. 2017; 8(42): 71965– 80. 29069761 PMC5641104 sourcePartly supported

  The Discussion (page unavailable) reports that a different study found increased tumor immunity after Abcg1 deletion in young mice on a Western-type diet, but not normal chow, and protection in aged mice on chow. Our copy does not report a genetic model of hypercholesterolemia as another condition in that study; it also says the present study observed protection without the diet or age requirement. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [76] Sag D, Cekic C, Wu R, Linden J, and Hedrick CC. The cholesterol transporter ABCG1 links cholesterol homeostasis and tumour immunity. Nat Commun. 2015; 6: 6354. 25724068 PMC4347884 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

Motifs are learnt as convolutional filters that are parameters of the model.

“Motifs are learnt as convolutional filters that are parameters of the model.”

• cites [46] Bhogale S, Seward C, Stubbs L, and Sinha S. SEAMoD: A fully interpretable neural network for cis-regulatory analysis of differentially expressed genes. bioRxiv. 2023. sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

The SEAMoD tool uses an interpretable neural network model to relate cis-regulatory sequences (promoters and enhancers) to gene expression in varying cellular contexts.

“This tool uses an interpretable neural network model to relate cis-regulatory sequences (promoters and enhancers) to gene expression in varying cellular contexts.”

• cites [46] Bhogale S, Seward C, Stubbs L, and Sinha S. SEAMoD: A fully interpretable neural network for cis-regulatory analysis of differentially expressed genes. bioRxiv. 2023. sourceSupported

May combine two claims.

Elevated expression of ABCA1 in tumors is associated with increased recurrence free survival when considering all breast cancer subtypes collectively, or Luminal A, Luminal B or Basal subtypes separately.

“Importantly, elevated expression of ABCA1 in tumors is associated with increased recurrence free survival, when considering all breast cancer subtypes collectively, or when only considering Luminal A, Luminal B or Basal subtypes separately (Fig. 1D).”

May combine two claims.

Multiplex immunofluorescence of human breast tumors found that while the majority of cells stained positive for ABCA1, CD11B+ myeloid cells displayed gradients of expression, generally higher than other cell types.

“Multiplex immunofluorescence (mIF) of human breast tumors found that while the majority of cells stained positive for ABCA1, CD11B+ myeloid cells displayed gradients of expression, generally higher than other cell types (Fig. 1A).”

May combine two claims.

siRNA-mediated knockdown of ABCA1 resulted in decreased ABCA1 mRNA, protein and cholesterol efflux.

“siRNA-mediated knockdown of ABCA1 resulted in the expected decrease in ABCA1 mRNA, protein and cholesterol efflux (Supplementary Fig. 1C–G).”

May combine two claims.

The effects on T cell expansion were restricted to CD8+ T cells as CD4+ T cell expansion was not significantly affected.

“Interestingly, the effects on T cell expansion were restricted to CD8+ T cells as CD4+ T cell expansion was not significantly affected (Supplementary Fig. 4C).”

May combine two claims.

siRNA knockdown of ABCA1 in BMDMs resulted in decreased T cell migration towards cancer cells, as did treatment of BMDMs with PSC833.

“siRNA knockdown of ABCA1 in BMDMs resulted in decreased T cell migration towards cancer cells, as did treatment of BMDMs with PSC833 (Fig. 5A, right panel).”

May combine two claims.

A correlation was found between the percentage of CD11B+ cells also expressing ABCA1 and the percentage of CD8+ cells; tumors with the highest three quartiles of CD11B+, ABCA1+ double positive cells were associated with the highest percentage of CD8+ cells.

“Likewise, a correlation was found between the percentage of CD11B+ cells also expressing ABCA1 and the percentage of CD8+ cells (Supplementary Fig. 5C & E); tumors with the highest three quartiles of CD11B+, ABCA1+ double positive cells were associated with the highest percentage of CD8+ cells (Fig. 6L).”

May combine two claims.

scRNA-seq of human breast tumors reveals elevated expression of ABCA1 within myeloid cells, and in particular macrophages, consistent across three different cohorts.

“scRNA-seq of human breast tumors also reveals elevated expression of ABCA1 within myeloid cells, and in particular macrophages; consistent across three different cohorts (Fig. 1C).”

May combine two claims.

siRNA knockdown of ABCA1 increased efferocytosis of apoptotic E0771 cancer cells, as did PSC833.

“More relevant to cancer biology, we also found that siRNA knockdown of ABCA1 increased efferocytosis of apoptotic E0771 cancer cells, as did PSC833 (Fig. 4H–I).”

May combine two claims.

With respect to the PI3K-AKT pathway, phospho-AKT1S1 and TSC2 were observed, but AKT2 was decreased.

“For example, with respect to the PI3K-AKT pathway, we observed phospho- AKT1S1 and TSC2, but a decrease in AKT2.”

May combine two claims.

Although several aspects of angiogenesis are upregulated in macrophages transfected with siRNA against ABCA1, they are not transitioned to a clear M2-state, at least as determined by gene expression.

“However, although several aspects of angiogenesis are upregulated in macrophages transfected with siRNA against ABCA1, they are not transitioned to a clear M2-state, at least as determined by gene expression (Fig. 7).”

May combine two claims.

Different AKT isoforms are involved, through both PI3K dependent and independent mechanisms.

“Mechanistically, different AKT isoforms are involved, through both PI3K dependent and independent mechanisms.”

May combine two claims.

In human breast tumors, ABCA1 expression was positively correlated with CD8 T cell abundance as well as enzymes indicative of CD8 T cell activity (IFNγ, GZMB and PRF1).

“In human breast tumors, ABCA1 expression was positively correlated with CD8 T cell abundance as well as enzymes indicative of CD8 T cell activity (IFNγ, GZMB and PRF1; Fig. 6).”

May combine two claims.

A PI3K inhibitor did not attenuate the effects of siABCA1 on efferocytosis, whereas PI3K inhibition did reduce the effects of siABCA1 on T cell expansion and cytotoxic functions.

“However, a PI3K inhibitor did not attenuate the effects of siABCA1 on efferocytosis, whereas PI3K inhibition did reduce the effects of siABCA1 on T cell expansion and cytotoxic functions.”

May combine two claims.

These robust effects appear to require direct contact between macrophages and T cells, although some enhancement was observed when the cell types were isolated by a membrane.

“These robust effects appear to require direct contact between macrophages and T cells, although some enhancement was observed when the cell types were isolated by a membrane (Fig. 5, Supplementary Fig. 4).”

May combine two claims.

The resulting expanded CD8+ T cells had less or more anti-cancer cytotoxic capacity when cultured with macrophages lacking or overexpressing ABCA1 respectively.

“Furthermore, the resulting expanded CD8+ T cells had less or more anti-cancer cytotoxic capacity when cultured with macrophages lacking or overexpressing ABCA1 respectively.”

May combine two claims.

Elevated ABCA1 is associated with improved response to ICB, decreased VEGFA angiogenesis, and increased CD8+ cell expansion and infiltration in tumors.

“These data are strongly supported by correlational data in humans, demonstrating that elevated ABCA1 is associated with (1) improved response to ICB, (2) decreased VEGFA angiogenesis, and (3) increased CD8+ cell expansion and infiltration in tumors.”

May combine two claims.

Loss of ABCA1 in macrophages impairs CD8+ T cell expansion while overexpression enhances it.

“We find that loss of ABCA1 in macrophages impairs CD8+ T cell expansion while overexpression enhances it.”

May combine two claims.

Findings in murine models that ABCA1 within myeloid cells alter CD8 T cell expansion and activity are supported by data from human tumors, suggesting conserved biology between the two species and that this is a relevant axis in humans.

“Therefore, our findings in murine models that ABCA1 within myeloid cells alter CD8 T cell expansion and activity are supported by data from human tumors, suggesting conserved biology between the two species and that this is a relevant axis in humans.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The study did not have sufficient power to make conclusions regarding HER2+ tumors.

“We did not have sufficient power to make conclusions regarding HER2+ tumors.”

May combine two claims.

Classic macrophage polarization states of M1 or M2 were not significantly enriched, suggesting that loss of ABCA1 re-educated macrophages into unique functional states.

“Classic macrophage polarization states of M1 or M2 were not significantly enriched for, suggesting that loss of ABCA1 re-educated macrophages into unique functional states.”

May combine two claims.

Some as yet unknown factor downstream of ABCA1 activity influences neo-angiogenesis, potentially through a decrease in VEGFA.

“Therefore, some as yet unknown factor downstream of ABCA1 activity influences neo-angiogenesis, potentially through a decrease in VEGFA.”

• AssumptionRests on an assumption: Promotion of angiogenesis is a key attribute of tumor associated macrophages and is generally seen as pro-tumor.

May combine two claims.

ABCA1 does not influence total phagocytic activity but does decrease efferocytosis, so there is likely a net increase in immunogenic phagocytosis.

“Therefore, ABCA1 does not influence total phagocytic activity but does decrease efferocytosis, so there is likely a net increase in immunogenic phagocytosis (non-efferocytotic phagocytosis).”

Probably not a substantial claim: may not pass S3 (load-bearing).

The analysis was underpowered to make firm conclusions regarding associations in HER2+ cases.

“Our analysis was underpowered to make firm conclusions regarding associations in HER2+ cases.”

Probably not a substantial claim: may not pass S2 (checkable).

Modulating ABCA1 activity within MCs may represent a novel approach to immune therapy.

“Therefore, modulating ABCA1 activity within MCs may represent a novel approach to immune therapy.”

May combine two claims.

AKT is a common feature governing the effects of T cell expansion/function and efferocytosis, but the specific subtypes differ, as does the dependence on upstream PI3K.

“Thus, while AKT is a common feature governing the effects of T cell expansion/function and efferocytosis, the specific subtypes differ, as does the dependence on the upstream PI3K.”

May combine two claims.

Myeloid cell ABCA1 activity supports CD8 T cell expansion and function, and this is likely clinically relevant and positions ABCA1 as a potential therapeutic target.

“Thus, our data indicating that myeloid cell ABCA1 activity supports CD8 T cell expansion and function are likely clinically relevant, and position ABCA1 as a potential therapeutic target.”

May combine two claims.

ABCA1 shifts macrophages into an alternate polarity, subtly enhancing several aspects of their anti-tumor capabilities.

“This reinforces our model whereby ABCA1 shifts macrophages into an alternate polarity, subtly enhancing several aspects of their anti-tumor capabilities.”

May combine two claims.

ABCA1 activity has critical implications in the regulation of various myeloid cell functions, including tumor infiltration, efferocytosis, and the support of T cell expansion and anti-cancer cytotoxic activities.

“We demonstrate that the activity of the cholesterol efflux and translocation protein, ABCA1, has critical implications in the regulation of various myeloid cell functions, including tumor infiltration, efferocytosis, and the support of T cell expansion and anti-cancer cytotoxic activities.”

May combine two claims.

ABCA1 and ABCG1 are not redundant, and may even have opposing effects.

“These data provide more support that ABCA1 and ABCG1 are not redundant, and may even have opposing effects.”

May combine two claims.

Loss of ABCA1 results in increased membrane cholesterol and hence lipid rafts and associated PI3K activity.

“Thus, we propose a model whereby loss of ABCA1 results in increased membrane cholesterol and hence lipid rafts and associated PI3K activity.”

Probably not a substantial claim: may not pass S2 (checkable).

ABCA1 represents an upstream node, ripe for therapeutic exploitation.

“Thus, by regulating several different anti-tumor associated functions in myeloid cells, ABCA1 represents an upstream node, ripe for therapeutic exploitation.”