Programmed cell removal by calreticulin in tissue homeostasis and cancer

Feng, Mingye; Marjon, Kristopher D.; Zhu, Fangfang; Weissman-Tsukamoto, Rachel; Levett, Aaron; Sullivan, Katie L.; Kao, Kevin S.; Markovic, Maxim; Bump, Paul; Jackson, Hannah M.; Choi, Timothy; Chen, Jing; Banuelos, Allison; Liu, Jie; Gip, Phung; Cheng, Lei; Wang, Guoyin; Weissman, Irving L.

doi:10.1038/s41467-018-05211-7

Cited by 127 publications

(120 citation statements)

References 68 publications

(93 reference statements)

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“…Because CALR can induce dendritic cell maturation, increasing antigen internalization and presentation, the biological function is beneficial for CALR in tumor growth control. A body of ligands were involved in facilitating CALR-mediated phagocytosis activity, such as C1q, CD91, mannose binding lectin (MBL) (18,19), S-nitrosothiol (SNO)-surfactant protein D (SNO-SP-D) (20,21), asialoglycans (22), lipoprotein receptor-related protein (LRP) (23), scavenger receptor A (SRA) (24), adiponectin and phosphatidylserine (25,26). After interaction with these potential ligands, CALR is phosphorylated (27) and subsequently activates endocytic receptor protein CD91 or called alpha-2-macroglobulin receptor on macrophages, finally triggering some downstream signaling pathways, such as p38 MAPK (mitogen-activated protein kinase) and NF-kappaB (20,28).…”

Section: Introductionmentioning

confidence: 99%

Calreticulin Blockade Attenuates Murine Acute Lung Injury by Inducing Polarization of M2 Subtype Macrophages

et al. 2020

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Section: Introductionmentioning

confidence: 99%

Calreticulin Blockade Attenuates Murine Acute Lung Injury by Inducing Polarization of M2 Subtype Macrophages

et al. 2020

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“…Under normal conditions, cell homeostasis is maintained by the balance of pro-and anti-phagocytic signals. Calreticulin is a dominant pro-phagocytic signal (commonly known as an 'eat me' signal) that is upregulated on the surfaces of cancer cells as well as damaged and aged cells, indicating to the phagocytic system that the cells should be eliminated 47 . However, due to high levels of CD47, which dominates the pro-phagocytic calreticulin signal, these cells fail to be eliminated 20 .…”

Section: Upregulation Of the Pro-phagocytic Signal Calreticulin In 16mentioning

confidence: 99%

“…However, due to high levels of CD47, which dominates the pro-phagocytic calreticulin signal, these cells fail to be eliminated 20 . Importantly, translocation of calreticulin to the cell surface of target cells determines cell removal 47 . Here, we assessed cell surface levels of calreticulin (CRT) by flow cytometry in the differentiated NPCs and OPCs.…”

Section: Upregulation Of the Pro-phagocytic Signal Calreticulin In 16mentioning

confidence: 99%

“…Furthermore, CRT binds to asialoglycans on the surface of cells, therefore the more potential binding sites, desialated glycans, the more potential calreticulin there is to promote interrogation and clearance by the phagocytic system. The absence of sialic acid on glycans can be determined by using a lectin, Phaseolus Vulgaris Leucoagglutinin (PHA-L) 47 . Staining with PHA-L on NPCs and OPCs derived from control and 16p_CNV iPSCs shows a marked increase in PHA-L binding in 16p_del subjects ( Figure 5C and 5F), indicating increased presence of asialoglycan binding sites for CRT.…”

Section: Upregulation Of the Pro-phagocytic Signal Calreticulin In 16mentioning

confidence: 99%

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Overexpression of CD47 is associated with brain overgrowth in 16p11.2 deletion syndrome

Brickler

Banuelos

et al. 2019

Preprint

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One of the most common genetic linkages associated with neuropsychiatric disorders, such as autism spectrum disorder and schizophrenia, occurs at the 16p11.2 locus. Copy number variants (CNVs) of the 16p gene can manifest in opposing head sizes. 16p11.2 deletion carriers tend to have macrocephaly (or brain enlargement), while those with 16p11.2 duplication frequently have microcephaly. Increases in both gray and white matter volume have been observed in brain imaging studies in 16p11.2 deletion carriers with macrocephaly. Here, we use human induced pluripotent stem cells (hiPSCs) derived from controls and subjects with 16p11.2 deletion and 16p11.2 duplication to understand the underlying mechanisms regulating brain overgrowth. To model both gray and white matter, we differentiated patient-derived iPSCs into neural progenitor cells (NPCs) and oligodendrocyte progenitor cells (OPCs). In both NPCs and OPCs, we show that CD47 (a 'don't eat me' signal) is overexpressed in the 16p11.2 deletion carriers contributing to reduced phagocytosis both in vitro and in vivo. Treatment of 16p11.2 deletion NPCs and OPCs with an anti-CD47 antibody to block CD47 restores phagocytosis to control levels. Furthermore, 16p11.2 deletion NPCs and OPCs upregulate cell surface expression of calreticulin (a pro-phagocytic 'eat me' signal) and its binding sites, indicating that these cells should be phagocytosed but fail to be eliminated due to elevations in CD47. While the CD47 pathway is commonly implicated in cancer progression, we document a novel role for CD47 in regulating brain overgrowth in psychiatric disorders and identify new targets for therapeutic intervention.

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“…Increased tumor cell surface expression of calreticulin can also occur (Kroemer et al, 2013). Calreticulin is a major prophagocytic signal that promotes phagocytosis by macrophages (Chao et al, 2010b;Feng et al, 2018).…”

Section: Immunogenic Cell Deathmentioning

confidence: 99%

Harnessing the Tumor Microenvironment for the Treatment of Double Hit Lymphoma

Lossos

Liu

Christie

et al. 2016

Blood

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Double Hit Lymphomas (DHL) constitute 6-9% of all newly-diagnosed Diffuse Large B-cell Lymphomas (DLBCL) and are defined by MYC translocation in concert with BCL2 (80%) or BCL6 (20%) rearrangement. DHL is characterized by poor upfront response to standard chemoimmunotherapies. The development of more effective treatment for DHL has been limited by the lack of models that accurately recapitulate the biology of this disease. To faithfully model bona fide human DHL, we generated two patient-derived xenografts (PDXs) in NSG mice from patients with DHL who relapsed after or were refractory to R-CHOP. Both PDXs had low/negative expression of surface CD20, in line with previous reports of DHL, and high surface expression of CD52. Importantly, these features recapitulated the original tumor phenotypes. We reasoned that immunologic approaches that target DHLs may help overcome their resistance to cytotoxic chemotherapy. Given the surface expression of CD52 on both PDXs, we treated xenografted mice with the anti-CD52 antibody Alemtuzumab (2x5mg/kg) when >2% tumor was detected in peripheral blood, a time corresponding to marked splenomegaly and ~30% bone marrow involvement. In both models, Alemtuzumab markedly reduced tumor involvement in the peripheral blood and spleen; however, the bone marrow was largely refractory to Alemtuzumab treatment. This is in line with previous results (Pallasch et al. Cell 2014) who also observed bone marrow-specific resistance to Alemtuzumab using a humanized transgenic model of DHL. In the latter model, Alemtuzumab resistance was successfully overcome by high-dose cyclophosphamide (CTX, 100mg/kg), which induced a tumor secretory response that induced macrophage-mediated phagocytosis of tumor cells. Thus, we treated both PDX models with vehicle, CTX 100mg/kg, Alemtuzumab or CTX+Alemtuzumab. While CTX monotherapy was able to partially reduce bone marrow tumor burden, in both PDXs, a single dose of CTX+Alemtuzumab was highly synergistic and eradicated nearly all bone marrow disease (Figure 1), which resulted in significantly extend murine survival compared to either CTX or Alemtuzumab alone alone (p<0.01). In contrast, high-dose Doxorubicin (DOX, 5mg/kg) had minimal effect on bone marrow DHL and DOX+Alemtuzumab was not synergistic. Strikingly, CD11b+F4/80+CD11c- macrophages comprised 20-40% of bone marrow cellularity 8 days after CTX treatment of xengorafted mice. DHL cells isolated from treated mice were next examined for changes in surface protein expression. At 16 hours after treatment, bone marrow DHL cells in mice receiving CTX (but not those receiving DOX or vehicle) had significantly reduced levels of surface CD47, the “don't eat me” signal that suppresses macrophage phagocytosis. By 48 hours after treatment, bone marrow DHL cells in mice receiving CTX (but not those receiving DOX or vehicle) had significantly increased expression of the pro-phagocytic proteins CD31 and Calreticulin. Next, we performed cytokine array analysis on bone marrow from mice engrafted with DHL PDXs at 16 hours after treatment. CTX-treated (but not vehicle- or DOX-treated) bone marrow had significantly increased levels of human IL-16 and CCL4, cytokines known to promote monocyte recruitment, as well as VEGF, PDGF and TNFα. Ex-vivo phagocytosis assays using murine bone marrow-derived macrophages confirmed that recombinant human VEGF and TNFα each enhanced macrophage phagocytosis of Alemtuzumab coated Raji lymphoma cells. Findings from RNA-seq studies on tumor cells and macrophages from the spleen and bone marrow as well as therapy induced changes in these respective niches will be further discussed at the meeting. In summary, we have demonstrated that high-dose CTX induces a phenotypic and secretory response from human DHL cells in vivo that promotes phagocytosis of Alemtuzumab-opsinized cells. As a result, we are initiating a clinical trial of this combination in patients with relapsed/refractory DHL. Figure 1. Percent tumor cells isolated from bone marrow of mice xenografted with DHL #61614 (left) or #63014 (right) on day 8 after treatment with cyclophosphamide (CTX), alemtuzumab (Alem) or the combination. Each marker indicates a mouse. Figure 1. Percent tumor cells isolated from bone marrow of mice xenografted with DHL #61614 (left) or #63014 (right) on day 8 after treatment with cyclophosphamide (CTX), alemtuzumab (Alem) or the combination. Each marker indicates a mouse. Disclosures No relevant conflicts of interest to declare.

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Programmed cell removal by calreticulin in tissue homeostasis and cancer

Cited by 127 publications

References 68 publications

Calreticulin Blockade Attenuates Murine Acute Lung Injury by Inducing Polarization of M2 Subtype Macrophages

Calreticulin Blockade Attenuates Murine Acute Lung Injury by Inducing Polarization of M2 Subtype Macrophages

Overexpression of CD47 is associated with brain overgrowth in 16p11.2 deletion syndrome

Harnessing the Tumor Microenvironment for the Treatment of Double Hit Lymphoma

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