Levels Of Colony-stimulating Factor-1 Research Articles

Colony stimulating factor-1 (CSF-1) signalling is predictive of response to immune checkpoint inhibitors in advanced non-small cell lung cancer

The identification of biomarkers related to treatment in patients with non-small cell lung cancer (NSCLC) treated with immune checkpoint inhibitors (ICIs) represents a significant challenge. The aim of this study was to determine the predictive value of macrophage-related markers assessed in plasma and tissue samples of patients with NSCLC undergoing ICI treatment. This bicentric study included a prospective cohort of 88 patients with advanced NSCLC who received first-line therapy with ICI (either as monotherapy or in combination with chemotherapy) or chemotherapy alone (CT). Samples were collected from the patients at baseline and during follow-up. Plasma levels of CSF-1 and IL-34 were measured using ELISA, while expression levels of the macrophage receptors CD163 and CSF-1-R were evaluated using immunohistochemistry on lung biopsies. At baseline, the median plasma CSF-1 expression was higher in patients who did not respond to immunotherapy compared to those who responded (8898 pg/mL vs. 14031 pg/mL, p = 0.0005). Importantly, high CSF-1 levels at the initial assessment were associated with disease progression regardless of the treatment received. Furthermore, high CSF-1 levels were associated with shorter progression-free survival (PFS) and overall survival (OS) in patients receiving ICI therapy, but not in those treated with chemotherapy. There was no correlation between IL-34, CSF-1R, CD163 and therapeutic response. We observed in vitro that the activation of lymphocytes mediated by pembrolizumab was hindered by the treatment of PBMC with recombinant CSF-1, suggesting that CSF-1 creates a systemic immunosuppressive state that interferes with ICI treatment. In conclusion, baseline CSF-1 levels represent a potential predictive marker to ICI treatment in NSCLC.

Validation of CSF-1 receptor (CD115) staining for analysis of murine monocytes by flow cytometry.

CD115, the receptor for colony stimulating factor 1, is essential for survival and differentiation of monocytes and macrophages and is therefore frequently used to define monocyte subsets and their progenitors in immunological assays. However, CD115 surface expression and detection by flow cytometry is greatly influenced by cell isolation and processing methods, organ source, and disease context. In a systematic analysis of murine monocytes, we define experimental conditions that preserve or limit CD115 surface expression and staining by flow cytometry. We also find that, independent of conditions, CD115 surface levels are consistently lower in Ly6Clo monocytes than in Ly6Chi monocytes, with the exception of Ly6Clo monocytes in the bone marrow. Furthermore, in contrast to IL-34, the presence of colony stimulating factor 1 impairs CD115 antibody staining in a dose-dependent manner, which, in a model of ischemic kidney injury with elevated levels of colony stimulating factor 1, influenced quantification of kidney monocytes. Thus, staining and experimental conditions affect quantitative and qualitative analysis of monocytes and may influence experimental conclusions.

Inhibition of inflammation and infiltration of M2 macrophages in NSCLC through the ATF3/CSF1 axis: Role of miR-27a-3p.

Non-small cell lung cancer (NSCLC) imposes a significant economic burden on patients and society due to its low overall cure and survival rates. Tumour-associated macrophages (TAM) affect tumour development and may be a novel therapeutic target for cancer. We collected NSCLC and tumour-adjacent tissue samples. Compared with the tumour-adjacent tissues, the Activation Transcription Factor 3 (ATF3) and Colony Stimulating Factor 1 (CSF-1) were increased in NSCLC tissues. Levels of ATF3 and CSF-1 were identified in different cell lines (HBE, A549, SPC-A-1, NCI-H1299 and NCI-H1795). Overexpression of ATF3 in A549 cells increased the expression of CD68, CD206 and CSF-1. Moreover, levels of CD206, CD163, IL-10 and TGF-β increased when A549 cells were co-cultured with M0 macrophages under the stimulation of CSF-1. Using the starbase online software prediction and dual-luciferase assays, we identified the targeting between miR-27a-3p and ATF3. Levels of ATF3, CSF-1, CD206, CD163, IL-10 and TGF-β decreased in the miR-27a mimics, and the tumour growth was slowed in the miR-27a mimics compared with the mimics NC group. Overall, the study suggested that miR-27a-3p might inhibit the ATF3/CFS1 axis, regulate the M2 polarization of macrophages and ultimately hinder the progress of NSCLC. This research might provide a new therapeutic strategy for NSCLC.

CSF-1 and Notch signaling cooperate in macrophage instruction and tissue repair during peripheral limb ischemia.

Ischemia causes an inflammatory response featuring monocyte-derived macrophages (MF) involved in angiogenesis and tissue repair. Angiogenesis and ischemic macrophage differentiation are regulated by Notch signaling via Notch ligand Delta-like 1 (Dll1). Colony stimulating factor 1 (CSF-1) is an essential MF lineage factor, but its role in ischemic macrophage development and the interaction with Notch signaling is so far unclear. Using a mouse model of hind limb ischemia with CSF-1 inhibitor studies and Dll1 heterozygous mice we show that CSF-1 is induced in the ischemic niche by a subpopulation of stromal cells expressing podoplanin, which was paralleled by the development of ischemic macrophages. Inhibition of CSF-1 signaling with small molecules or blocking antibodies impaired macrophage differentiation but prolonged the inflammatory response, resulting in impaired perfusion recovery and tissue regeneration. Yet, despite high levels of CSF-1, macrophage maturation and perfusion recovery were impaired in mice with Dll1 haploinsufficiency, while inflammation was exaggerated. In vitro, CSF-1 was not sufficient to induce full MF differentiation from donor monocytes in the absence of recombinant DLL1, while the presence of DLL1 in a dose-dependent manner stimulated MF differentiation in combination with CSF-1. Thus, CSF-1 is an ischemic niche factor that cooperates with Notch signaling in a non-redundant fashion to instruct macrophage cell fate and maturation, which is required for ischemic perfusion recovery and tissue repair.

Gene Expression Levels of CSF-1 and CSF-1R Endometrial under The Influence of Prolactin Level in Unexplained Miscarriage: A Case-Control Study.

Hormones such as prolactin, by influencing expression of the endometrial genes, play a pivotal role in embryo implantation and development. The present study aimed to evaluate serum level of prolactin and its effect on altering expression level of colony-stimulating factor-1 (CSF-1) and colony-stimulating factor-1 receptor (CSF-1R) genes in endometrial tissue during in vitro fertilization (IVF) pregnancy in the infertile women and recurrent pregnancy loss (RPL), compared to fertile women, who lost their pregnancies at gestational age <20 weeks. In this case-control study, 40 infertile women, 40 IVF pregnant women with RPL and 40 fertile women who lost their pregnancies at <20 weeks of gestation for unknown reasons were selected. Prolactin serum level was assessed using ELISA technique and expression of CSF-1 and CSF-1R genes was determined in endometrial tissue, using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Mean prolactin level of the infertile group was 24.38 ± 1.43 ng/mL and it had statistically significant relationship with the fertile group (P<0.001). Expression level of the CSF-1 and CSF-1R genes were higher in the fertile than infertile groups by 2.88 times (P<0.0001) and 2.64 times (P<0.0001), while it was respectively 2.28 (P<0.0001) and 1.69 (P<0.0001) times higher compared to the RPL group. Risk factors for pregnancy loss, such as aging, increased body mass index (BMI), smoking and diabetes caused decreasing changes in gene expression (CSF-1 and CSF-1R ) and the differences were statistically significant, except in the infertile group. The present study showed a significant relationship of CSF-1 and CSF-1R expression levels with pregnancy loss. Risk factors such as aging, obesity, smoking and diabetes decreased both genes expression levels.

Blockade of tumor-derived colony-stimulating factor 1 (CSF1) promotes an immune-permissive tumor microenvironment

The macrophage colony-stimulating factor 1 (CSF1) is a chemokine essential for the survival, proliferation, and differentiation of mononuclear phagocytes from hemopoietic stem cells. In addition to its essential physiological role in normal tissues, the CSF1/CSF1 receptor axis is known to be overexpressed in many tumor types and associated with poor prognosis. High levels of CSF1 within the tumor microenvironment have been shown to recruit and reeducate macrophages to produce factors that promote tumor invasiveness and accelerate metastasis. In this study, we demonstrate, for the first time, that treating established syngeneic murine colon and breast carcinoma tumors with a CSF1R-blocking antibody also promotes the expansion of neoepitope-specific T cells. To assess the role of tumor-derived CSF1 in these model systems, we generated and characterized CSF1 CRISPR-Cas9 knockouts. Eliminating tumor-derived CSF1 results in decreased tumor growth and enhanced immunity against tumor-associated neoepitopes, potentially promoting an immune permissive tumor microenvironment in tumor-bearing mice. The combination of neoepitope vaccine with anti-PDL1 in the MC38 CSF1−/− tumor model significantly decreased tumor growth in vivo. Moreover, anti-CSF1R therapy combined with the adeno-TWIST1 vaccine resulted in tumor control, decreased metastasis, and a synergistic increase in CD8 T cell infiltration in 4T1 mammary tumors. Analysis of the tumor microenvironment demonstrated greater CD8 T cell infiltration and a reduction in tumor-associated macrophages following CSF1R inhibition in both tumor models. Our findings thus add to the therapeutic potential of CSF1 targeting agents by employing combinations with vaccines to modulate anti-neoepitope responses in the tumor microenvironment.

First-in-human phase 1 study of pimicotinib (ABSK021), a CSF-1R inhibitor, in patients with advanced solid tumors.

3100 Background: Colony-stimulating factor 1 (CSF-1) pathway is involved in the development of various types of cancer. Pimicotinib is an orally available, selective, potent small molecule CSF-1R inhibitor with significant pre-clinical anti-tumor activity. Here we present the results of the phase 1 trial. Methods: The escalation part employed a 3+3 design with a starting dose of 25 mg QD (28-day/ cycle) to determine the maximum tolerated dose (MTD) and the recommended dose for expansion (RDE). Additional pts with selected tumor types including pancreatic carcinoma, triple-negative breast cancer, lung cancer, and sarcoma were treated at RDE in the expansion part for further evaluation. Tenosynovial giant cell tumor (TGCT) cohorts were analyzed and reported separately. Results: As of December 31, 2022, 74 pts received at least one dose of pimicotinib at 25 mg (n=6), 50 mg (n=58), 75 mg (n=8), or 100 mg (n=2) QD. respectively. Median age was 59 y; 68.9% were female; 47.3% were white, 33.8% were Asian and 18.9% were African American. 74.3% were ECOG PS 1; 85.1% had metastasis; 67.6% had ≥3 lines of prior systemic anti-tumor therapies. Median duration of treatment was 49.5 days (range 4-377). Treatment-related adverse events (TRAEs), Grade ≥3 TRAEs, and serious TRAEs occurred in 74.3%, 21.6%, and 4.1% of pts, respectively. The most common TRAEs (≥20%) were serum enzyme elevations, including CPK, AST and LDH, which were all believed to be pimicotinib MOA related, with CPK increase (14.9%) being the only Grade ≥3 TRAE reported in ≥10% of pts. 3 pts (4.1%) reported serious TRAEs of ascites, blood bilirubin increase, and vaginal hemorrhage. After single and multiple dosing, pimicotinib plasma exposure increased slightly less than dose proportion. The rapid oral absorption (median tmax of 0.87 - 1.52 hrs) with moderate terminal t1/2 (mean t1/2 ranges: 43.6 - 63.5 hrs) suggested sustained drug exposure was achieved with a once daily dosing schedule. No dose adjustment was needed based on tested covariates including weight, age and race. Pimicotinib treatment led to a more than 85% reduction of non-classical monocytes (NCM) across all dose levels. Plasma CSF-1 levels were significantly increased from 25mg QD to 50mg QD and reached a plateau at 50mg QD, with ~40-fold maximal induction. Reduced CD163+ macrophages in skin and tumor tissues were seen at 50mg QD. All tested doses showed adequate inhibition of NCM, and CSF-1 change showed a positive correlation with plasma pimicotinib concentration. The MTD and RDE were determined as 75 mg QD and 50 mg QD. Conclusions: Tolerable safety profile and favorable PK/PD properties supported pimicotinib to be further evaluated in advanced solid tumors, including combinations. Clinical trial information: NCT04192344 .

Determining the expression levels of CSF-1 and OCT4, CREM-1, and protamine in testicular biopsies of adult Klinefelter patients: Their possible correlation with spermatogenesis.

Klinefelter syndrome (KS) is the most prevalent genetic disorder of infertile males. This study aimed to determine in Klinefelter patients (KS) the expression levels of spermatogenic markers and testicular growth factors that might predict spermatogenesis based on conventional testicular sperm extraction (TESE). The expression levels of the pre-meiotic (OCT4, CD9, GFR-α1, α-6-INTEGRIN, SALL4, C-KIT), meiotic (CREM-1), and post-meiotic (protamine) markers, as well as the colony stimulating factor-1 (CSF-1) were examined in testicular biopsies with and without mature sperm of KS and normal karyotype of azoospermic patients (AZO) with complete spermatogenesis. In the biopsies of AZO, the expression levels (fold of expression compared to the PPI of the same sample) of OCT4 were 9.68± 7.93, CREM 42.78± 28.22, CSF-1 3.07 ± 3.19, and protamine 78498.12 ± 73214.40. Biopsies from KS included 7 with sperm and 17 without sperm. Among the biopsies with sperm, the expression levels of OCT4 were 7.27± 9.29, CREM 3.13± 7.89, CSF-1 35.5 ± 48.01, and protamine 902.97 ± 2365.92. In 14 biopsies without sperm, we found low expression levels of OCT4, CREM and CSF-1, and no expression of protamine. However, in three of the biopsies without sperm that highly expressed OCT4 and CSF-1, the expression levels of CREM-1 and protamine were high. These results may be used for further consulting with patients considering repeating conventional TESE or micro TESE and cryopreservation for possible future in-vitro spermatogenesis.

Spinal Cord Stimulation Alleviates Neuropathic Pain by Attenuating Microglial Activation via Reducing Colony-Stimulating Factor 1 Levels in the Spinal Cord in a Rat Model of Chronic Constriction Injury.

BACKGROUND:Spinal cord stimulation (SCS) is an emerging, minimally invasive procedure used to treat patients with intractable chronic pain conditions. Although several signaling pathways have been proposed to account for SCS-mediated pain relief, the precise mechanisms remain poorly understood. Recent evidence reveals that injured sensory neuron-derived colony-stimulating factor 1 (CSF1) induces microglial activation in the spinal cord, contributing to the development of neuropathic pain (NP). Here, we tested the hypothesis that SCS relieves pain in a rat model of chronic constriction injury (CCI) by attenuating microglial activation via blocking CSF1 to the spinal cord.METHODS:Sprague-Dawley rats underwent sciatic nerve ligation to induce CCI and were implanted with an epidural SCS lead. SCS was delivered 6 hours per day for 5 days. Some rats received a once-daily intrathecal injection of CSF1 for 3 days during SCS.RESULTS:Compared with naive rats, CCI rats had a marked decrease in the mechanical withdrawal threshold of the paw, along with increased microglial activation and augmented CSF1 levels in the spinal dorsal horn and dorsal root ganglion, as measured by immunofluorescence or Western blotting. SCS significantly increased the mechanical withdrawal threshold and attenuated microglial activation in the spinal dorsal horn in CCI rats, which were associated with reductions in CSF1 levels in the spinal dorsal horn and dorsal roots but not dorsal root ganglion. Moreover, intrathecal injection of CSF1 completely abolished SCS-induced changes in the mechanical withdrawal threshold and activation of microglia in the spinal dorsal horn in CCI rats.CONCLUSIONS:SCS reduces microglial activation in the spinal cord and alleviates chronic NP, at least in part by inhibiting the release of CSF1 from the dorsal root ganglion ipsilateral to nerve injury.

Gingival crevicular fluid CSF-1 and IL-34 levels in patients with stage III grade C periodontitis and uncontrolled type 2 diabetes mellitus.

Periodontal diseases are inflammatory conditions that alter the host's response to microbial pathogens. Type 2 diabetes mellitus (T2DM) is a complex disease that affects the incidence and severity of periodontal diseases. This study investigated the gingival crevicular fluid (GCF) levels of colony-stimulating factor-1 (CSF-1) and interleukin-34 (IL-34) in patients with stage III grade C periodontitis (SIII-GC-P) and stage III grade C periodontitis with uncontrolled type 2 diabetes (SIII-GC-PD). In total, 72 individuals, including 24 periodontally healthy (PH), 24 SIII-GC-P, and 24 SIII-GC-PD patients, were recruited for this study. Periodontitis patients (stage III) had interdental attachment loss (AL) of 5 mm or more, probing depth (PD) of 6 mm or more, radiographic bone loss advancing to the middle or apical part of the root, and tooth loss (<5) due to periodontal disease. Radiographic bone loss in the teeth was also evaluated; grade C periodontitis was defined as a ratio of the percentage of root bone loss to age greater than 1.0. The plaque index (PI), gingival index (GI), presence of bleeding on probing (BOP), PD, and clinical AL were used for clinical periodontal assessments. GCF samples were obtained and analyzed using an enzyme-linked immunosorbent assay. All clinical parameters-PD, AL, GI, BOP, and PI-were significantly higher in the SIII-GC-PD group than in the PH and SIII-GC-P groups for both the full mouth and each sampling site (P<0.05). The total IL-34 and CSF-1 levels were significantly higher in the SIII-GC-PD group than in the PH and SIII-GC-P groups (P<0.05), and there were significant differences between the periodontitis groups (P<0.05). These findings suggest that IL-34 and CSF-1 expression increases in patients with SIII-GC-PD. CSF-1 was associated with the inflammatory status of periodontal tissues and T2DM, while IL-34 was associated only with T2DM. ClinicalTrials.gov Identifier: NCT04891627.

CircCDC45 promotes the malignant progression of glioblastoma by modulating the miR-485-5p/CSF-1 axis

BackgroundGlioblastoma (GBM) is characterized by progressive growth and metastasis. Numerous studies claim that the deregulation of circular RNAs (circRNAs) is associated with cancer progression. However, the role of circRNAs in GBM is largely limited. The purpose of this study was to investigate the functions of circCDC45 in GBM and provide a feasible functional mechanism to support its role.MethodsThe expression of circCDC45, miR-485-5p and colony-stimulating factor 1 (CSF-1) mRNA was examined using quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was assessed using cell counting kit − 8 (CCK-8) assay and colony formation assay. Cell migration and cell invasion were monitored using transwell assay. The protein levels of proliferation-related markers and CSF-1 were determined using western blot. The target relationship was predicted using bioinformatics tools and validated using dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Animal models were constructed to verify the role of circCDC45 in vivo.ResultsThe expression of circCDC45 and CSF-1 was elevated in GBM tissues and cells, while the expression of miR-485-5p was declined. Downregulation of circCDC45 or CSF-1 blocked GBM cell proliferation, invasion and migration as well as tumor growth in vivo. In mechanism, circCDC45 positively regulated the expression of CSF-1 by targeting miR-485-5p. Inhibition of miR-485-5p reversed the biological effects caused by circCDC45 downregulation in GBM cells.ConclusionCircCDC45 promoted the progression of GBM by mediating the miR-485-5p/CSF-1 axis, and circCDC45 might be a promising plasmatic biomarker for GBM diagnosis and treatment.

Levels of myeloid‐related proteins in saliva for screening and monitoring of periodontal disease

To evaluate the salivary levels of myeloid-related markers in relation to periodontal disease and their potential screening capability, as well as the effects of periodontal treatment on these markers in periodontitis patients. Participants with a healthy periodontium (n=60) and with gingivitis (n=63) and periodontitis (n=72) were recruited. Periodontitis patients received non-surgical treatment and were re-examined after 3 and 6months. Unstimulated saliva was collected at baseline and at 1, 3, and 6 months after therapy for the periodontitis patients. Levels of colony-stimulating factor-1 (CSF-1), interleukin-34 (IL-34), S100A8/A9, S100A12, hepatocyte growth factor (HGF), IL-1β, and matrix metalloproteinase-8 (MMP-8) were analysed by immunoassays. CSF-1, S100A8/A9, S100A12, IL-1β, MMP-8, and HGF were significantly elevated in saliva from periodontitis and gingivitis patients in comparison to healthy individuals, whereas IL-34 was significantly lower in periodontitis compared to both healthy individuals and gingivitis patients. IL-34 increased significantly 3 months after treatment, while IL-1β and MMP-8 decreased 1 month after therapy. Additionally, periodontitis patients clustered in high and low levels of S100A8/A9, whereby those with high levels had more bleeding, deeper pockets, and higher S100A12. Salivary levels of myeloid-related markers are altered in periodontitis and are partially modulated by periodontal treatment. Measuring S100A8/A9 in saliva may identify distinct groups of periodontitis patients.

Targeting the CSF1/CSF1R axis is a potential treatment strategy for malignant meningiomas.

Malignant meningiomas are fatal and lack effective therapy. As M2 macrophages are the most prevalent immune cell type in human meningiomas, we hypothesized that normalizing this immunosuppressive population would be an effective treatment strategy. We used CIBERSORTx to examine the proportions of 22 immune subsets in human meningiomas. We targeted the colony-stimulating factor 1 (CSF1) or CSF1 receptor (CSF1R) axis, an important regulator of macrophage phenotype, using monoclonal antibodies (mAbs) in a novel immunocompetent murine model (MGS1) for malignant meningioma. RNA sequencing (RNA-seq) was performed to identify changes in gene expression in the tumor microenvironment (TME). Mass cytometry was used to delineate changes in immune subsets after treatment. We measured patients' plasma CSF1 levels using ELISA and CSF1R expression using multiplex quantitative immunofluorescence in a human meningioma tissue microarray. Human meningiomas are heavily enriched for immunosuppressive myeloid cells. MGS1 recapitulates the TME of human meningiomas, including an abundance of myeloid cells, a paucity of infiltrating T cells, and low programmed death ligand 1 (PD-L1) expression. Treatment of murine meningiomas with anti-CSF1/CSF1R, but not programmed cell death receptor 1 (PD-1), mAbs abrogate tumor growth. RNA-seq and mass cytometry analyses reveal a myeloid cell reprogramming with limited effect on T cells in the TME. CSF1 plasma levels are significantly elevated in human patients, and CSF1R is highly expressed on CD163+ macrophages within the human TME. Our findings suggest that anti-CSF1/CSF1R antibody treatment may be an effective normalization cancer immunotherapy for malignant meningiomas.

The Presence of Colony-Stimulating Factor-1 and Its Receptor in Different Cells of the Testis; It Involved in the Development of Spermatogenesis In Vitro.

Spermatogenesis is a complex process, in which spermatogonial cells proliferate and differentiate in the seminiferous tubules of the testis to generate sperm. This process is under the regulation of endocrine and testicular paracrine/autocrine factors. In the present study, we demonstrated that colony stimulating factor-1 (CSF-1) is produced by mouse testicular macrophages, Leydig, Sertoli, peritubular cells and spermatogonial cells (such as CDH1-positively stained cells; a marker of spermatogonial cells). In addition, we demonstrated the presence of CSF-1 and its receptor (CSF-1R) in testicular macrophages, Leydig, Sertoli, peritubular cells and spermatogonial cells of human testis. We also show that the protein levels of CSF-1 were the highest in testis of 1-week-old mice and significantly decreased with age (2–12-week-old). However, the transcriptome levels of CSF-1 significantly increased in 2–3-week-old compared to 1-week-old, and thereafter significantly decreased with age. On the other hand, the transcriptome levels of CSF-1R was significantly higher in mouse testicular tissue of all examined ages (2–12-week-old) compared to 1-week-old. Our results demonstrate the involvement of CSF-1 in the induction the proliferation and differentiation of spermatogonial cells to meiotic and postmeiotic stages (BOULE- and ACROSIN-positive cells) under in vitro culture conditions, using methylcellulose culture system (MCS). Thus, it is possible to suggest that CSF-1 system, as a testicular paracrine/autocrine system, is involved in the development of different stages of spermatogenesis and may be used in the development of future therapeutic strategies for treatment of male infertility.

Abstract 2020: Identification of abnormal CSF1 transcripts in tenosynovial giant cell tumors and dose-dependent increase in plasma CSF1 levels in response to pexidartinib treatment

Abstract Background: Tenosynovial giant cell tumors (TGCTs) are characterized by rearrangements of the colony-stimulating factor 1 (CSF1) gene. Dysregulated CSF1 may attract CSF1 receptor (CSF1R)-bearing mononuclear cells that form the bulk of the tumor. CSF1R inhibitors including pexidartinib have been developed and have proven to be effective therapies. Methods: Formalin-fixed paraffin-embedded (FFPE) TGCT specimens from the phase 1 first-in-human study of pexidartinib (NCT01004861) were analyzed using a custom sequencing assay to detect CSF1 genetic alterations. The ArcherDX RNA panel (ArcherDX, Inc., Boulder, CO) consisted of 33 gene-specific primer sets specifically designed to target CSF1, including at least 1 primer set to target each exon/exon boundary as well as primer sets to tile the 3′-untranslated region (3′UTR). Plasma samples from 5 phase 1 trials of pexidartinib across multiple tumor types (NCT02777710, NCT01217229, NCT01525602, NCT01790503, NCT02452424) were assayed for CSF1 protein by solid-phase ELISA (Quantikine® Human MCSF, R&amp;D Systems, Inc., Minneapolis, MN) at baseline and following pexidartinib (200-1200 mg). Results: FFPE TGCT specimens (N=25) were successfully isolated, prepared into libraries, and sequenced; 8 showed evidence of gene rearrangements at the junction of CSF1 exons 5/6, and 15 showed alterations of the CSF1 3′UTR (n=23). All 25 TGCT libraries had higher CSF1 expression, with 24 exceeding 2-fold of the RNA control library. The plasma samples from 132 patients were analyzed for CSF1 protein. Pexidartinib treatment (200-1200 mg) led to a dose-dependent increase in plasma CSF1 by day 8, 15, or 29. Significant (&amp;gt;4-fold) CSF1 elevation was observed at 600 mg or higher doses. Discussion: All 25 analyzed TGCT study patients had tumor tissue with elevated CSF1 ligand transcripts, and 23 had CSF1 genomic alterations identified, thus confirming the TGCT etiology for which pexidartinib therapy was recently approved. The initial discovery of gross chromosomal aberrations involving the CSF1 locus used break-apart FISH probes (West et al. Proc Natl Acad Sci USA. 2006;103:690-695). Details of the CSF1 gene aberrations were obtained here using a sequencing assay suitable for FFPE specimens having partially degraded RNA. Abnormal CSF1 transcripts identified in our study are similar to those published by others (Ho et al. Genes Chromosomes Cancer. 2019[Epub]; Tsuda et al. Int J Cancer. 2019;145:3276-3284) after completion of our work, including both predicted fusion proteins and loss of 3′UTR microRNA negative regulatory sites. Increased plasma CSF1 is a useful pharmacodynamics marker of CSF1R inhibition. One potential mechanism for CSF1 plasma elevations could be the inhibition of liver macrophages, known to express CSF1R and thought to play an essential role in the normal clearance of CSF1 from circulation. Citation Format: Paul Severson, Brian L. West, William D. Tap, Zev A. Wainberg, Sandra Tong-Starksen, Henry H. Hsu, Chao Zhang. Identification of abnormal CSF1 transcripts in tenosynovial giant cell tumors and dose-dependent increase in plasma CSF1 levels in response to pexidartinib treatment [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2020.

Reticular Fibroblasts Expressing the Transcription Factor WT1 Define a Stromal Niche that Maintains and Replenishes Splenic Red Pulp Macrophages

Located within red pulp cords, splenic red pulp macrophages (RPMs) are constantly exposed to the blood flow, clearing senescent red blood cells (RBCs) and recycling iron from hemoglobin. Here, we studied the mechanisms underlying RPM homeostasis, focusing on the involvement of stromal cells as these cells perform anchoring and nurturing macrophage niche functions in lymph nodes and liver. Microscopy revealed that RPMs are embedded in a reticular meshwork of red pulp fibroblasts characterized by the expression of the transcription factor Wilms' Tumor 1 (WT1) and colony stimulating factor 1 (CSF1). Conditional deletion of Csf1 in WT1+ red pulp fibroblasts, but not white pulp fibroblasts, drastically altered the RPM network without altering circulating CSF1 levels. Upon RPM depletion, red pulp fibroblasts transiently produced the monocyte chemoattractants CCL2 and CCL7, thereby contributing to the replenishment of the RPM network. Thus, red pulp fibroblasts anchor and nurture RPM, a function likely conserved in humans.

Synergistic alveolar bone resorption by diabetic advanced glycation end products and mechanical forces.

The association between diabetes mellitus (DM) and bone diseases is acknowledged. However, the mechanistic pathways leading to the alveolar bone (AB) destruction remain unclear. This study aims to elucidate the mechanical forces (MF)-induced AB destruction in DM and its underlying mechanism. In vivo periodontal tissue responses to MF were evaluated in rats with diabetes. In vitro human periodontal ligament (PDL) cells were either treated with advanced glycation end products (AGEs) alone or with AGEs and MF. In vivo, the transcription of VEGF-A, colony stimulating factor-1 (CSF-1), and Ager was upregulated in diabetes, whereas changes in DDOST and Glo1 mRNAs were negligible. DM induced VEGF-A protein in the vascular cells of the PDL and subsequent angiogenesis, but DM itself did not induce osteoclastogenesis. MF-induced AB resorption was augmented in DM, and such augmentation was morphologically substantiated by the occasional undermining resorption as well as the frontal resorption of the AB by osteoclasts. The mRNA levels of CSF-1 and vascular endothelial growth factor (VEGF) during MF application were highly elevated in diabetes, compared with those of the normal counterparts. In vitro, AGEs treatment elevated Glut-1 and CSF-1 mRNA levels via the p38 and JNK pathways, whereas OGT and VEGF levels remained unchanged. Compressive MF especially caused upregulation of VEGF, CSF-1, and Glut-1 levels, and such upregulation was further enhanced by AGEs treatment. Overloaded MF and AGEs metabolites may synergistically aggravate AB destruction by upregulating CSF-1 and VEGF. Therefore, regulating the compressive overloading of teeth, as well as the levels of diabetic AGEs, may prove to be an effective therapeutic modality for managing DM-induced AB destruction.

Acetaminophen reduces apical root resorption during orthodontic tooth movement in rats

ObjectiveThe present study aimed to investigate the inhibitory effect of acetaminophen on apical root resorption during orthodontic tooth movement by controlling inflammation in the periodontal ligament and apical pulp tissue. MethodsHuman periodontal ligament and pulp cells were subjected to 10 kPa of cyclic tensile force (CTF) in a Flexcell Strain Unit for 48 h. Then, 10 and 100 μM acetaminophen were added to the culture medium, and the expression of interleukin (IL)-1B, receptor activator of nuclear factor kappa-B ligand (RANKL), tumor necrosis factor (TNF)α, and colony stimulating factor 1 (CSF1) were evaluated. In an animal experiment, the upper first molars of 7-week-old rats were moved mesially by applying 10 g of orthodontic force. After 30 days of force application, the effects of acetaminophen on apical root resorption were examined. ResultsIn both the periodontal ligament and pulp cells, the expression levels of IL-1B, TNFα, RANKL, and CSF1 were significantly higher in the CTF-treated group than in the control group. However, the expression levels of these factors were decreased by acetaminophen administration. High expression of IL-1B, TNFα, RANKL, and CSF1 at the root apex were also detected immunohistochemically in rats after tooth movement, but were decreased by acetaminophen administration. In addition, the number of odontoclasts and the amount of apical root resorption were significantly decreased in the acetaminophen group. Importantly, no significant difference in tooth movement was observed between the acetaminophen and control groups. ConclusionsThese results suggest that acetaminophen can reduce severe root resorption in the apex area without disturbing orthodontic tooth movement.

Increased expression of colony‐stimulating factor‐1 in mouse spinal cord with experimental autoimmune encephalomyelitis correlates with microglial activation and neuronal loss

Microglia contribute to pathophysiology at all stages of multiple sclerosis. Colony-stimulating factor-1 (CSF1) is crucial for microglial proliferation and activation. In this study we measured the CSF1 levels and studied its cellular expression in the mouse spinal cords with experimental autoimmune encephalomyelitis (EAE) to explore the potential contribution of CSF1 in neuronal death. ELISA data showed that CSF1 levels were significantly higher in the spinal cords with acute and chronic EAE than those of normal and adjuvant-injected mice. Immunohistochemical studies demonstrated that CSF1 was expressed in astrocytes and neurons in normal mouse spinal cord. In acute EAE, CSF1 expression was significantly increased, especially in astrocytes in peripheral white matter and large motoneurons. High density of activated microglia was observed in the gray matter where motoneurons expressed high-level CSF1 in acute EAE. Significant large motoneuron loss was seen in chronic EAE and the remaining motoneurons with high-level CSF1 were enwrapped by microglia. Viral vector mediated over-expression of CSF1 in spinal neurons induced profound proliferation and activation of microglia at the injection site and microglia enwrapped CSF1-transduced neurons and their neurites. Significant loss of large CSF1-transduced neurons was seen at 2 and 3 weeks post-viral injection. Demyelination in the CSF1-transduced areas was also significant. These results implicate that CSF1 upregulation in CNS may play an important role in the proliferation and activation of microglia in EAE, contributing to neuroinflammation and neurodegeneration. © 2018 Wiley Periodicals, Inc.

CSF-1 and Ang-2 serum levels — prognostic and diagnostic partners in non-small cell lung cancer

BackgroundLung cancer is the most incident and lethal form of cancer, with late diagnosis as a major determinant of its bad prognosis. Immunotherapies targeting immune checkpoints improve survival, but positive results encompass only 30%–40% of the patients, possibly due to alternative pathways to immunosuppression, including tumour-associated macrophages (TAM). Colony stimulating factor-1 (CSF-1) is implicated in TAM differentiation and recruitment to tumours and in tumour angiogenesis, through a special setting of Tie-2-expressing macrophages, which respond to angiopoietin-2 (Ang-2). We evaluated the role of serum levels of CSF-1 in non-small cell lung cancer (NSCLC) prognosis and whether these could serve as biomarkers for NSCLC detection, along with Ang-2.Participants and methodsWe prospectively studied an unselected cohort of 145 patients with NSCLC and a group of 30 control individuals. Serum levels of Ang-2 and CSF-1 were measured by ELISA prior to treatment.ResultsSerum levels of CSF-1 and Ang-2 are positively correlated (p<0.000001). Individuals with high serum levels of CSF-1 have a 17-fold risk for NSCLC presence and patients with combined High Ang-2/CSF-1 serum levels present a 5-fold increased risk of having NSCLC. High Ang-2/CSF-1 phenotype is also associated with worst prognosis in NSCLC.ConclusionsCombined expression of CSF-1 and Ang-2 seems to contribute to worst prognosis in NSCLC and it is worthy to understand the basis of this unexplored partnership. Moreover, we think CSF-1 could be included as a biomarker in NSCLC screening protocols that can improve the positive predictive value of the current screening modalities, increase overall cost effectiveness and potentially improve lung cancer survival.