Colony-stimulating Factor 1 Receptor Research Articles

Abstract LB077: CSF-1R inhibition with Pimicotinib (ABSK021) enhanced anti-tumor efficacy of KRASG12C inhibitors in preclinical non-small cell lung cancer mouse models

Abstract Background: KRAS mutations are highly prevalent oncogenic drivers in human cancers and KRASG12C is the most frequent mutations found in KRAS-mutant non-small lung cancer (NSCLC). It is reported that tumor associated macrophages (TAMs) are enriched in KRAS-driven transgenic lung cancer models. Depletion of macrophage results in a significant decrease in tumor burden and increased survival. Sotorasib (AMG510), the first KRASG12C inhibitor approved by FDA, has been found to modulate tumor immunity, impacting the recruitment of macrophages. CSF-1R signaling is critical for macrophage proliferation and survival. CSF-1R inhibition depletes TAMs and reprograms tumor microenvironment (TME), resulting in enhancement of anti-tumor immunity. Therefore, the combination of KRASG12C and CSF-1R inhibitors may synergize to enhance anti-tumor efficacy. Pimicotinib (ABSK021) is a potential best-in-class small molecule inhibitor of CSF-1R in clinical development of multiple indications. Here, we conducted in vivo experiments to explore the combined effects of KRASG12C and CSF-1R inhibitors. Methods: In vivo efficacy studies were performed to evaluate the combined treatment of KRASG12C inhibitors (AMG510 and ABSK071, a next-generation inhibitor targeting KRASG12C with better inhibitory activity) and CSF-1R inhibitor (ABSK021) in NCI-H2122 PBMC humanized mice and LLC syngeneic mice. RNA sequencing was employed to investigate the mechanism of action underlying the synergistic anti-tumor effect. Immunohistochemistry (IHC) was utilized to examine the changes of immune cell populations in TME. Results: The combinatory regimen of ABSK021 and KRASG12C inhibitors showed improved anti-tumor effects than each single agent alone in both mouse NSCLC models. Bioinformatics and follow-up analysis revealed a reduction in immune suppressive TAMs in tumors treated with ABSK021 alone or in combination with KRASG12C inhibitors. Furthermore, modulation of TME related to anti-tumor immunity, such as activated CD4 T, CD8 T and NK cells, was increased, and pro-tumor immunity, such as MDSC, regulatory T cells, was decreased in combination group. IHC results confirmed that ABSK021 addition led to macrophage depletion and more CD8 T cell infiltration in TME. Conclusions: For the first time, we demonstrate that the combined inhibition of KRASG12C and CSF-1R leads to superior therapeutic efficacy in preclinical NSCLC mouse models. This results suggests a potential novel therapeutic regimen that could yield improved clinical benefit to patients. Citation Format: Nannan Zhang, Bin Shen, Shenyan Liu, Cheng Dai, Haiyan Ying, Fushen Guo, Zhixuan Zhu, Wan Shi, Min Tu, Jie Wang, Jie Zhang, Manqi Liu, Qi Zhang, Zhui Chen. CSF-1R inhibition with Pimicotinib (ABSK021) enhanced anti-tumor efficacy of KRASG12C inhibitors in preclinical non-small cell lung cancer mouse models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB077.

Abstract CT266: Initial efficacy of surufatinib plus sintilimab and IBI310 for patients with high-grade advanced-neuroendocrine neoplasm: A multicenter, single arm phase 2 stud

Abstract Background: High-grade advanced-neuroendocrine neoplasm (HG-NEN) are more aggressive and have worse prognosis. Surufatinib (SUR, a selective VEGFRs; FGFR1; and CSF-1R inhibitor). Ipilimumab (anti-CTLA-4) plus nivolumab (anti-PD-1) has been recommended for extrapulmonary and non-pancreatic NEC if the disease progresses following chemotherapy by the NCCN guidelines for neuroendocrine tumors. Preclinical and clinical studies have shown that SUR is a good partner of immune checkpoint inhibitors (ICIs). In this study, we present findings of SUR combined with Sintilimab (SIN, anti-PD-1) and IBI310 (IBI, anti-CTLA-4) for HG-NEN patients (pts). Methods: Pts with histologically confirmed HG-NENs who resistant or intolerant to standard treatment were included in the study. Enrolled pts received SUR (250 mg, qd, po, q3w) combined with SIN (200 mg, d1, i.v.gtt, q3w) and IBI (1 mg/kg, d1, i.v.gtt, q6w). The primary endpoint was ORR per RECIST 1.1. The second endpoints were PFS, DCR, safety and OS. Results: By Dec 29, 2023, 14 pts were enrolled and available for efficacy assessment. Median age, 53 yrs (range 22- 66); male, 71.4%; ECOG PS 1, 50%; GEP-NEN, 57.1%; NEC, 71.4%; Median ki-67 index, 65% (range 25%-90%); liver metastases, 57.1%. 85.7% pts received prior systemic therapy (50% 1L and 35.7% ≥2L). Confirmed ORR was 28.6% and DCR 76.8%. Median PFS was 4.15 months. Among 9 pts with ≤1 prior systemic regimens, confirmed ORR and DCR were 44.4% and 88.9%, respectively, with a median PFS of 4.5 months. Out of 14 pts, all experienced some form of treatment- emergent events (TEAEs), with only 3 participants (21.4%) reporting grade 3 TEAEs. These included manageable events like increased bilirubin, immune pancreatitis, gastric retention, and increased amylase, each with a 7.1% incidence. No grade 4 TEAEs or new safety signals occurred. Conclusion: Surufatinib combined with Sintilimab and IBI310 showed a potential antitumor activity and a tolerable safety profile, providing an additional treatment option for pts with high-grade NEN. Citation Format: Panpan Zhang, Yakun Wang, Jie Li, Lin Shen, Ming Lu. Initial efficacy of surufatinib plus sintilimab and IBI310 for patients with high-grade advanced-neuroendocrine neoplasm: A multicenter, single arm phase 2 stud [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT266.

Abstract CT134: Safety and immunologic impact of neoadjuvant/adjuvant GM-CSF-secreting allogenic pancreatic tumor cell vaccine (GVAX) combined with cyclophosphamide, pembrolizumab, and macrophage-targeting CSF1R inhibitor IMC-CS4 in pancreatic adenocarcinoma

Abstract Background: Strategies are needed to improve effectiveness of immunotherapy in pancreatic ductal adenocarcinoma (PDA). Our recently published study (NCT02648282) showed that increased M1/M2 ratio and decreased PDL1+ M2-like tumor associated macrophages (TAM) in locally advanced PDA is associated with longer overall survival (OS) following combination of stereotactic body radiotherapy, GVAX and anti-PD1 antibody pembrolizumab (Pem). We hypothesized targeting M2-like macrophages, regulated by the colony stimulating factor-1 (CSF1) pathway, would improve outcomes. In this pilot study we investigated the safety and intratumoral immune response of neoadjuvant/adjuvant triplet immunotherapy: GVAX/cyclophosphamide (CY), Pem, and CSF1 receptor blockade (IMC-CS4) in patients with resectable or borderline resectable PDA. Methods: After completing chemotherapy and radiation patients received two 3-week cycles of neoadjuvant triplet immunotherapy. Following surgery, patients received 4 additional cycles followed by booster Pem every 3 weeks and GVAX/CY every 6 months, for one year. Co-primary endpoints were safety, assessed by NCI Common Terminology Criteria for Adverse Events (AEs), and immune response (increased CD8+ T-cell density in surgical specimen as compared to pre-treatment core). Secondary endpoints included disease free survival (DFS), OS, surgical resectability, and pathologic response (PR). Results: Nine patients were enrolled. Two immune related grade 3/4 AEs (diarrhea and rash) were reported. Median DFS was 12.6 months (95% CI: 7.3, NR) and median OS was 20.4 months (95% CI: 17.5, NR). All patients underwent surgery with major PR rate of 78% (defined as <10% residual viable tumor), including one complete PR. Comparing paired biopsies, the study met its primary immunologic endpoint showing 6/8 patients (75%) had a >80% increase of CD8+ T cells; the increase was at least 1.8 times the baseline median absolute deviation following triple therapy (p=0.012). There was a significant increase of Granzyme B+ CD8+ T cells among all patients (p=0.045). No significant change in myeloid cell density, including TAM, was observed, suggesting IMC-CS4 did not deplete macrophages, but potentially reprogrammed them. The 4 patients with greatest drop in CD66b+ granulocyte levels had major PR (grade 0 or 1). Conclusion: The triplet was safe and significantly increased activated CD8+ T cells suggesting the combination can induce an effector T cell response. Decreased granulocyte levels correlated with PR. Citation Format: Arielle Urman, Yingjun Ding, Hao Wang, Richard A. Burkhart, Jin He, Elizabeth M. Jaffee, Amol K. Narang, Dung Le, Daniel A. Laheru, Lei Zheng, Ana De Jesus-Acosta. Safety and immunologic impact of neoadjuvant/adjuvant GM-CSF-secreting allogenic pancreatic tumor cell vaccine (GVAX) combined with cyclophosphamide, pembrolizumab, and macrophage-targeting CSF1R inhibitor IMC-CS4 in pancreatic adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT134.

Abstract CT094: Targeting CSF1R with BLZ945 results in effective peripheral and tumor immune microenvironment modulation in advanced solid tumors and may be associated with limited efficacy in recurrent non-mesenchymal glioblastoma

Abstract Colony-stimulating factor 1 receptor (CSF-1R) signaling is involved in regulating monocyte differentiation, and the recruitment and survival of tumor-associated macrophages (TAMs). BLZ945 is a small molecule, highly selective, orally bioavailable, potent kinase inhibitor of CSF-1R that has shown the ability to reduce the recruitment of TAMs in animal models and promote infiltration of effector T cells into the tumor in preclinical models. Given the potential effects of BLZ945 on effector T-cell activity, a combination of BLZ945 with the programmed death 1 (PD-1) inhibitor spartalizumab was investigated in 1) a dose escalation phase I clinical trial with advanced solid tumor patients; and 2) an expansion phase II clinical trial in adult patients with relapsed/refractory glioblastoma (GBM) after standard of care and other lines of therapy (NCT02829723). The percentages of non-classical and intermediate monocytes in the PBMC fraction were decreased after treatment at multiple doses and schedules of treatment tested in the escalation phase. Screening and on-treatment paired tumor biopsy samples from the escalation phase were analyzed by RNA sequencing. Downregulated genes have been noted as macrophage-specific genes, suggesting that treatment with BLZ945 is downregulating macrophages in the tumor microenvironment. There appeared to be a trend for a reduction in TAM gene signatures and increase in T-cell inflammation signatures upon treatment with single-agent BLZ945 or BLZ945 + spartalizumab. RNA-sequencing analysis from archival surgical resection GBM samples from the expansion phase, revealed a potential correlation between PDGFRA gene expression and best percent change in tumor size. This suggests that non-mesenchymal GBM, for which PDGFRA is part of the defining signature, are more sensitive to the treatment. DNA-sequencing of these archival samples also revealed mutations in p53 in the tumors with partial responses but no mutations in EGFR or TERT. Citation Format: Carlos Rodrigo Gil Del Alcazar, Virginia Xu, Chia-chi Lin, Marta Gil-Martin, Aung Naing, Liqiong Fan, Fang Yang, David Quinn, Jincheng Wu, Cornelia Quadt, Jennifer Mataraza. Targeting CSF1R with BLZ945 results in effective peripheral and tumor immune microenvironment modulation in advanced solid tumors and may be associated with limited efficacy in recurrent non-mesenchymal glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT094.

Macrophage colony-stimulating factor and its role in the tumor microenvironment: novel therapeutic avenues and mechanistic insights.

The tumor microenvironment is a complex ecosystem where various cellular and molecular interactions shape the course of cancer progression. Macrophage colony-stimulating factor (M-CSF) plays a pivotal role in this context. This study delves into the biological properties and functions of M-CSF in regulating tumor-associated macrophages (TAMs) and its role in modulating host immune responses. Through the specific binding to its receptor colony-stimulating factor 1 receptor (CSF-1R), M-CSF orchestrates a cascade of downstream signaling pathways to modulate macrophage activation, polarization, and proliferation. Furthermore, M-CSF extends its influence to other immune cell populations, including dendritic cells. Notably, the heightened expression of M-CSF within the tumor microenvironment is often associated with dismal patient prognoses. Therefore, a comprehensive investigation into the roles of M-CSF in tumor growth advances our comprehension of tumor development mechanisms and unveils promising novel strategies and approaches for cancer treatment.

The plasma-soluble CSF1R level is a promising prognostic indicator for pediatric Langerhans cell histiocytosis.

Langerhans cell histiocytosis (LCH) is a rare hematologic neoplasm characterized by the clonal proliferation of Langerhans-like cells. Colony-stimulating factor 1 receptor (CSF1R) is a membrane-bound receptor that is highly expressed in LCH cells and tumor-associated macrophages. In this study, a soluble form of CSF1R protein (sCSF1R) was identified by plasma proteome profiling, and its role in evaluating LCH prognosis was explored. We prospectively measured plasma sCSF1R levels in 104 LCH patients and 10 healthy children using ELISA. Plasma sCSF1R levels were greater in LCH patients than in healthy controls (p<.001) and significantly differed among the three disease extents, with the highest level in MS RO+ LCH patients (p<.001). Accordingly, immunofluorescence showed the highest level of membrane-bound CSF1R in MS RO+ patients. Furthermore, the plasma sCSF1R concentration at diagnosis could efficiently predict the prognosis of LCH patients treated with standard first-line treatment (AUC=0.782, p<.001). Notably, dynamic monitoring of sCSF1R levels could predict relapse early in patients receiving BRAF inhibitor treatment. In vitro drug sensitivity data showed that sCSF1R increased resistance to Ara-C in THP-1 cells expressing ectopic BRAF-V600E. Overall, the plasma sCSF1R level at diagnosis and during follow-up is of great clinical importance in pediatric LCH patients.

Macrophage migration inhibitory factor blockade reprograms macrophages and disrupts prosurvival signaling in acute myeloid leukemia

The malignant microenvironment plays a major role in the development of resistance to therapies and the occurrence of relapses in acute myeloid leukemia (AML). We previously showed that interactions of AML blasts with bone marrow macrophages (MΦ) shift their polarization towards a protumoral (M2-like) phenotype, promoting drug resistance; we demonstrated that inhibiting the colony-stimulating factor-1 receptor (CSF1R) repolarizes MΦ towards an antitumoral (M1-like) phenotype and that other factors may be involved. We investigated here macrophage migration inhibitory factor (MIF) as a target in AML blast survival and protumoral interactions with MΦ. We show that pharmacologically inhibiting MIF secreted by AML blasts results in their apoptosis. However, this effect is abrogated when blasts are co-cultured in close contact with M2-like MΦ. We next demonstrate that pharmacological inhibition of MIF secreted by MΦ, in the presence of granulocyte macrophage-colony stimulating factor (GM-CSF), efficiently reprograms MΦ to an M1-like phenotype that triggers apoptosis of interacting blasts. Furthermore, contact with reprogrammed MΦ relieves blast resistance to venetoclax and midostaurin acquired in contact with CD163+ protumoral MΦ. Using intravital imaging in mice, we also show that treatment with MIF inhibitor 4-IPP and GM-CSF profoundly affects the tumor microenvironment in vivo: it strikingly inhibits tumor vasculature, reduces protumoral MΦ, and slows down leukemia progression. Thus, our data demonstrate that MIF plays a crucial role in AML MΦ M2-like protumoral phenotype that can be reversed by inhibiting its activity and suggest the therapeutic targeting of MIF as an avenue towards improved AML treatment outcomes.

Targeting the CSF-1/CSF-1R Axis: Exploring the Potential of CSF1R Inhibitors in Neurodegenerative Diseases.

We report herein the potential of colony-stimulating factor-1 receptor (CSF1R) inhibitors as therapeutic agents in neuroinflammatory diseases, with a focus on Alzheimer's disease (AD). Employing a carefully modified scaffold, N-(4-heterocycloalkyl-2-cycloalkylphenyl)-5-methylisoxazole-3-carboxamide, we identify highly selective and potent CSF1R inhibitors─7dri and 7dsi. Molecular docking studies shed light on the binding modes of these key compounds within the CSF1R binding site. Remarkably, kinome-wide selectivity assessment underscores the impressive specificity of 7dri for CSF-1R. Notably, 7dri emerges as a potent CSF-1R inhibitor with favorable cellular activity and minimal cytotoxicity among the synthesized compounds. Demonstrating efficacy in inhibiting CSF1R phosphorylation in microglial cells and successfully mitigating neuroinflammation in an in vivo LPS-induced model, 7dri establishes itself as a promising antineuroinflammatory agent.

Blocking Microglial Proliferation by CSF-1R Inhibitor Does Not Alter the Neuroprotective Effects of Adoptive Regulatory T Cells in 3xTg Alzheimer's Disease Mice.

Alzheimer's disease (AD) is a chronic neurodegenerative disease that causes cognitive impairment. Neuroinflammation induced by activated microglia exacerbates AD. Regulatory T cells (Tregs) play roles in limiting neuroinflammation by converting microglial polarization. Therefore, adoptive Treg therapy is considered an attractive option for neurodegenerative disorders. However, the mechanism underlying Treg therapy via microglial modulation is not fully understood. In this study, we sought to determine whether adoptively transferred Tregs were effective when microglia proliferation was inhibited by using GW2580, which is an inhibitor of CSF1R. We found that inhibition of microglial proliferation during Treg transfer did not alter the therapeutic effects of Tregs on cognitive deficits and the accumulation of Aβ and pTAU in 3xTg-AD mice. The expression of pro- and anti-inflammatory markers in the hippocampus of 3xTg mice showed that GW2580 did not affect the inhibition of neuroinflammation by Treg transfer. Additionally, adoptively transferred Tregs were commonly detected in the brain on day 7 after transfer and their levels decreased slowly over 100 days. Our findings suggest that adoptively transferred Tregs can survive longer than 100 days in the brain, suppressing microglial activation and thus alleviating AD pathology. The present study provides valuable evidence to support the prolonged efficacy of adoptive Treg therapy in AD.

Relative contributions of osteal macrophages and osteoclasts to postnatal bone development in CSF1R-deficient rats and phenotype rescue following wild-type bone marrow cell transfer.

Macrophage and osteoclast proliferation, differentiation and survival are regulated by colony-stimulating factor-1 receptor (CSF1R) signaling. Osteopetrosis associated with Csf1 and Csf1r mutations has been attributed to the loss of osteoclasts and deficiency in bone resorption. Here we demonstrate that homozygous Csf1r mutation in rat leads to delayed postnatal skeletal ossification associated with substantial loss of osteal macrophages (osteomacs) in addition to osteoclasts. Osteosclerosis and site-specific skeletal abnormalities were reversed by intraperitoneal transfer of wild-type bone marrow cells (BMT) at weaning. Following BMT, IBA1+ macrophages were detected before TRAP+ osteoclasts at sites of ossification restoration. These observations extend evidence that osteomacs independently contribute to bone anabolism and are required for normal postnatal bone growth and morphogenesis.

CSF1R inhibitor PLX3397 depletes microglia in Mongolian gerbil Meriones unguiculatus, but not in syrian hamster Mesocricetus auratus

Microglia are the residential immune cells in the central nervous system. Their roles as innate immune cells and regulators of synaptic remodeling are critical to the development and the maintenance of the brain. Numerous studies have depleted microglia to elucidate their involvement in healthy and pathological conditions. PLX3397, a blocker of colony stimulating factor 1 receptor (CSF1R), is widely used to deplete mouse microglia due to its non-invasiveness and convenience. Recently, other small rodents, including Syrian hamsters (Mesocricetus auratus) and Mongolian gerbils (Meriones unguiculatus), have been recognized as valuable animal models for studying brain functions and diseases. However, whether microglia depletion via PLX3397 is feasible in these species remains unclear. Here, we administered PLX3397 orally via food pellets to hamsters and gerbils. PLX3397 successfully depleted gerbil microglia but had no effect on microglial density in hamsters. Comparative analysis of the CSF1R amino acid sequence in different species hints that amino acid substitutions in the juxtamembrane domain may potentially contribute to the inefficacy of PLX3397 in hamsters.

Abstract 7161: Preclinical pharmacology modeling (pharmacokinetics/efficacy) of narazaciclib (HX301 or ON123300) to predict its clinical effective dose

Abstract Background: Acute myeloid leukemia (AML) is an aggressive leukemia of myeloid lineage. CSF1R (colony-stimulating factor 1 receptor) regulates the proliferation, differentiation and polarization of cells of myeloid lineages, e.g. monocytes/macrophages. CSF1R expression is aberrantly upregulated in many AML, leading to a hypothesis that CSF1R is a putative leukemogenic driver in a subset of AML and thus could be a potential target for treating AML. Previous studies demonstrated that Narazaciclib (HX301 or ON123300), an investigational kinase inhibitor (TKi), exhibited high potency activity against CSF1R (IC50 0.285 nM) and inhibition of macrophage proliferation and AML tumor growth both in vitro/in vivo, implicating its potential use as an AML therapeutic1 (An et al., 2023 AACR Annual Meeting). In addition, narazaciclib has recently completed the dose escalation phase 1 study in advanced solid tumors. However, the therapeutic window of narazaciclib as a CSF1Ri for treating AML remains to be determined. The present study attempted to utilize both preclinical PK and in vivo efficacy models to predict the effective clinical dose of narazaciclib for guiding the future AML clinical trials. Methods: A single dose of narazaciclib at various of dose levels was orally administrated in NOD/Scid mice, and plasma samples were collected at designated timepoints. The concentrations of narazaciclib in plasma were analyzed by LC-MS. In a Ba/F3-ETV6-CSF1R in vivo xenograft tumor model in Nod/Scid mice, narazaciclib at various dose levels was orally administrated daily. Tumor volumes were measured twice weekly. Results: The mouse PK study showed that narazaciclib displayed overall similar absorption pattern as seen in unpublished human clinical data. Narazaciclib was absorbed rapidly after oral dosing with a mean Tmax ranging from 1 to 2 hours in mouse. However, narazaciclib was cleared faster in mouse, with a mean T1/2 ranging from 1.9 to 2.8 hours, than in human. In addition, the exposures of narazaciclib, as measured by Cmax and AUC in plasma, increased in a relatively dose-proportional manner, as seen in human patients. In the Ba/F3-ETV6-CSF1R xenograft model where the tumor cell growth is solely dependent on the CSF1R kinase activity, narazaciclib at 10 mg/kg did not induce any anti-tumor effects with a TGI of -14.61%. In contrast, narazaciclib at 50 mg/kg and 100 mg/kg exerted significant anti-tumor effects with a TGI of 85.49% and 94.55%, respectively. Based on the mouse PK results, narazaciclib at 50 mg/kg in mouse is estimated to have an AUC of approximate 2000 h*ng/mL, which is equivalent to the dose of 160 - 200 mg/d in human patients. Therefore, 160 mg dose level is predicted to be an effective dose for narazaciclib in AML patients. Conclusions: Narazaciclib at 160 mg/d is predicted to be an effective dose in AML patients. Citation Format: Tao Yang, Hang Ke, Jinping Liu, Kefeng Gong, Henry Qixiang Li. Preclinical pharmacology modeling (pharmacokinetics/efficacy) of narazaciclib (HX301 or ON123300) to predict its clinical effective dose [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7161.

Abstract 1168: Anti-CSF1R antibody monotherapy inhibits minimal residual disease in a novel immunocompetent murine colorectal cancer metastasis model

Abstract Background: The high mortality rate of colorectal cancer (CRC) is primarily due to metastatic disease. Microscopic minimal residual disease (MRD) detected in patients by circulating tumor DNA (ctDNA) may remain radiographically undetectable and persist following therapeutic intervention, resulting in recurrence. Preclinical models are urgently needed to elucidate the mechanisms whereby MRD escapes anti-tumor immune surveillance. We have developed and characterized a preclinical liver MRD model that recapitulates immune response, with the goal to discover novel strategies to improve outcomes for patients with ctDNA+ MRD. Methods: We generated genetically engineered mouse colonic organoids CDX2 CRE; Rosa26 LSL-CAS9-GFP with APC and TP53 mutation and performed intrasplenic injection in syngeneic C57BL/6 mice to recapitulate microsatellite stable CRC metastasis to the liver. Mice were monitored for radiographically detectable disease using MRI. Tumors were resected at various timepoints, with comparison between metastases at various sizes of development (dichotomized as micro- and macro-metastases). We performed multiplex immunofluorescence staining, single cell RNA seq, and spatial transcriptomic analysis to delineate the immune compartments of micro- and macro-metastasis in untreated mice. After identifying colony stimulating factor -1 receptor (CSF1R) as a target, we performed a vehicle controlled anti-CSF1R monotherapy study for 4 weeks. Treatment began at either 7 days before metastasis is radiographically detectable to mimic treatment of MRD, or 21 days post tumor engraftment. Results: Immune exclusion was confirmed through intratumoral CD45+ leukocyte densities, with transition to an immune excluded phenotype occurring above 0.5mm in size (defined as macro-metastases). In our preclinical model, immunosuppressive populations of macrophages (IBA1+CD163+) dominated in the micro-metastases. This observation was confirmed in 13 patients with paired micro- and macro-metastases to the liver (p=0.0065). Strikingly, anti-CSF1R treatment in the murine model eradicated IBA1+CD163+ macrophages (p&amp;lt;0.0001) and induced complete remission of liver micro-metastasis (0/7 mice with residual disease), in contrast to limited efficacy for macro-metastases (5/5 mice with progressed radiographically disease). Additionally, a long-term study confirmed 100% survival (5/5 mice) and no evidence of disease via MRI at 6 months upon anti-CSF1R treatment compared to vehicle (0/5 mice). Conclusions: We established a reproducible preclinical model of MRD with similar immunopathological features of MRD in patients. Our data further identified infiltration of immunosuppressive macrophage populations as an early determinant of MRD progression and indicated CSF1R as a potential new clinical therapeutic target for ctDNA+-defined MRD patients. Citation Format: Alaa M. Mohamed, Kyung Serk Cho, Melinda Soeung, Amanda Anderson, Jumanah Alshenaifi, Ganiraju Manyam, Oscar Villareal, Jennifer Davis, Will Norton, Sisi Gao, Christopher Bristow, Federica Carbone, Stefania Napolitano, Oluwadara Coker, Isha Khanduri, Justin Huang, Dipen M. Maru, Mauro Di Pilato, Ryan Sun, Linghua Wang, David G. Menter, Giannicola Genovese, Giulio Draetta, Natalie Fowlkes, Scott Kopetz. Anti-CSF1R antibody monotherapy inhibits minimal residual disease in a novel immunocompetent murine colorectal cancer metastasis model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1168.

Abstract 1581: Colony-stimulating factor-1 receptor as a potential therapeutic target in pancreatic ductal adenocarcinoma

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) is associated with poor prognosis, rising incidence and suffers from limited existing treatment options. Currently, salient features of PDAC are being investigated that can potentially predict immunotherapy outcomes, to identify small patient populations that can benefit from such strategies. Preliminary studies show that colony-stimulating factor-1 receptor (CSF1R), a surface receptor tyrosine kinase that drives survival, function, proliferation and differentiation of several myeloid lineage cell types can be a potentially interesting target for combination immunotherapy. Methods: PDAC patients from the TCGA PanCancer Atlas were stratified into CSF1Rhi and CSF1Rlo cohorts and clinically relevant characteristics were compared between the two. The mRNA expression of CSF1R and immunological markers of interest were evaluated and Gene Set Enrichment Analysis (GSEA) was run on the two groups to analyze differential expression of gene sets. Results: mRNA expression z-scores were used to classify 129 patient samples in the TCGA PanCancer Atlas dataset into CSF1Rhi (z-score&amp;gt;1, n=20) and CSF1Rlo (z-score&amp;lt;0, n=109) cohorts. Both have similar disease staging, MSI MANTIS scores, tumor mutation burden, median diagnosis age, sex and race distribution. However, the CSF1Rhi cohort has a distinct, non-conventional PDAC driver gene mutation profile (genomic alteration frequencies in CSF1Rhi vs CSF1Rlo cohorts respectively: a) KRAS: 30% vs 73% b) CDKN2A: 15% vs 57% and c) SMAD4: 5% vs 42%) (p-value&amp;lt;0.01 for each). CSF1R correlates with markers of CD8+ T-cell function (R2 values: CD8A: 0.683, GZMB: 0.508, PRF1: 0.673), CD8+ T-cell infiltration (CXCL9: 0.596, CXCL10: 0.483, CCL5: 0.666) and expression of immune checkpoint genes (PDCD1: 0.602, CD274: 0.544, CTLA4: 0.613). CD8A, GZMB, PRF1, CXCL9, CXCL10, CCL5, PDCD1, CD274, CTLA4 are significantly upregulated in the CSF1Rhi group (log2Fold changes respectively: 2.07, 1.57, 1.48, 2.51, 1.71, 1.75, 1.63, 1.34, 1.80). GSEA against the hallmark gene set database shows that KRAS signaling, Inflammatory response, IL6 JAK STAT3 and Interferon-gamma signaling are significantly upregulated in the CSF1R high patient group (FDR q-value &amp;lt;0.01 for each). Conclusion: Preliminary in silico analysis suggests that PDAC patients with higher levels of CSF1R may be more immune infiltrated. Future studies will evaluate the potential of CSF1R as a biomarker to identify an immune infiltrated subset of PDAC patients ex vivo and the mechanism of immune infiltration in CSF1Rhi PDACs in vivo. Citation Format: Somak Chaudhuri, Dustin A. Deming, Cheri A. Pasch, Katherine A. Johnson, Chelsie K. Sievers, Philip B. Emmerich. Colony-stimulating factor-1 receptor as a potential therapeutic target in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1581.

Abstract 6648: Low-intensity pulsed ultrasound enhances the presentation of brain-derived antigens by facilitating the migration of antigen-presenting cells from the brain to the periphery

Abstract Gliomas, the most common primary brain tumors, remain largely incurable despite advancements in surgical and adjuvant therapies. While immunotherapies offer promising avenues, their efficacy has been limited by formidable anatomical barriers, such as the blood-brain barrier (BBB), which restricts the migration of immune cells into the brain and impedes the efficient drainage of brain-derived antigens, establishing an immune-privileged environment. Consequently, neoplastic formations within the brain often escape detection by the peripheral immune system, complicating therapeutic interventions. The application of low-intensity pulsed ultrasound with microbubbles (LIPU/MB) has emerged as a potential solution, enabling the transient opening of the BBB. This facilitates drug transport and immune cell trafficking, including Chimeric Antigen Receptor (CAR) T cells, into brain tumors. However, challenges such as antigen-loss escape persist with CAR T cells. One proposed solution involves promoting antigen/epitope spreading by inducing the presentation of brain (tumor) antigens to the systemic immune system. To investigate this, we conducted intravital imaging using Colony-stimulating factor receptor 1 (CSFR1) and CX3CR1 reporter mice, visualizing antigen-presenting cell locomotion in the brain before and after LIPU/MB treatment using two-photon microscopy. Within one hour after LIPU/MB, we observed the migration of CSFR1+ and CX3CR1+ immune cells from the brain parenchyma and perivascular space into the bloodstream, a phenomenon absent prior to LIPU/MB treatment. Beyond that, flow cytometry identified CD45intermediateCD11b+-CX3CR1+-TMEM119+-P2RY12+microglia-like' cells in the blood. To further assess the immunological effects of LIPU/MB on brain antigen presentation, we utilized transgenic mice expressing a minigene encoding four immunogenic peptides under the glial fibrillary acidic protein (GFAP) promoter. LIPU/MB treatment was associated with increased proliferation and priming of adoptively transferred CNS antigen-specific naïve T cells in cervical lymph nodes, ultimately homing into the CNS, indicating enhanced antigen presentation. Furthermore, in a subset of glioblastoma (GBM) patients undergoing LIPU/MB combined with doxorubicin and anti-PD-1 blocking antibodies, CD11b+CD14+P2YR12+ microglia were detected in blood samples during cycles 3-6 of treatment. This increase was absent in blood samples from healthy donors.Collectively, our findings suggest that LIPU/MB represents a novel approach to augment brain-antigen presentation in the periphery, holding potential implications for improving the immunotherapeutic management of gliomas.[M.G., V.A.A., A.M.S., and H.O. contributed equally to this work.] Citation Format: Marco Gallus, Victor Andres Arrieta, Atsuro Saijo, Pavlina Chuntova, Akane Yamamichi, Jeffrey Haegelin, Su Phyu, Lan Phung, Shayan A. Amir-Kabirian, Senthilnath Lakshmanachetty, Li Chen, Karl Habashy, Andrew Gould, Christina Amidei, Catalina Lee Chang, Roger Stupp, Michael Canney, Hideho Okada. Low-intensity pulsed ultrasound enhances the presentation of brain-derived antigens by facilitating the migration of antigen-presenting cells from the brain to the periphery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6648.

Abstract 2122: Enhanced anticancer efficacy via ROS-dependent ferroptosis: Synergy between surufatinib and cisplatin in small cell lung cancer

Abstract Objective: Small cell lung cancer (SCLC), a highly malignant subset of neuroendocrine carcinoma, accounts for 13% of lung cancer cases and is characterized by its high aggressiveness and poor prognosis. The longstanding first-line treatment, a platinum-etoposide chemotherapy regimen, is increasingly compromised by resistance issues, highlighting the critical need for new therapeutic approaches. Surufatinib, an inhibitor selectively targeting vascular endothelial growth factor receptors (VEGFR) 1, 2, and 3, fibroblast growth factor receptor type 1 (FGFR1), and colony-stimulating factor-1 receptor (CSF-1R), has demonstrated efficacy in a spectrum of solid tumors, including neuroendocrine neoplasm, thyroid cancers and biliary cancers. Recently, ferroptosis, an iron-dependent cell death pathway, offers therapeutic promise, especially by inhibiting the xCT system, effective in pancreatic and lung adenocarcinomas. Yet, surufatinib's potential for inducing ferroptosis in SCLC remains to be investigated. Methods: Cytotoxicity assays were performed on SCLC cell lines to assess the combinatorial effect of surufatinib and cisplatin, revealing a synergistic interaction. Ferroptosis inhibitors were applied to assess the reversal of cytotoxic effects. Mitochondrial morphology, reactive oxygen species (ROS) accumulation, membrane potential dissipation (JC-1 assay), lipid peroxidation (MDA levels), and intracellular iron accumulation (Fe2+ quantification) assays confirmed ferroptosis induction. Western blot analyses were conducted to explore the mechanistic pathways, supplemented by in vivo antitumor efficacy evaluations. Results: In vitro assays revealed that surufatinib, when combined with cisplatin, exerts a pronounced synergistic effect on small cell lung cancer (SCLC) cell lines by inducing ferroptosis. The combination therapy strategically modulated ferroptosis-related pathways: cisplatin effectively decreased the expression of glutathione peroxidase 4 (GPX4), critical for cellular antioxidant defenses, while surufatinib increased the expression of acyl-CoA synthetase long-chain family member 4 (ACSL4), which is essential for the accumulation of lipid peroxides. This dual pathway engagement significantly amplified the ferroptotic response, culminating in heightened antitumor activity. Corroborative in vivo experiments substantiated these findings, validating the combined treatment's efficacy in a living model and underscoring its potential as a robust therapeutic strategy for SCLC. Conclusion: Our findings reveal that surufatinib triggers ferroptosis in small cell lung cancer—a novel discovery. Additionally, it also unveils that surufatinib and cisplatin together enhance anti-cancer efficacy via the ROS-mediated ferroptosis pathway, marking a significant advance in SCLC therapy. Citation Format: Xiaolin Li, Ziting Zhang, Qiyun Tang, Huae Xu. Enhanced anticancer efficacy via ROS-dependent ferroptosis: Synergy between surufatinib and cisplatin in small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2122.

Bioactive materials from berberine-treated human bone marrow mesenchymal stem cells promote alveolar bone regeneration by regulating macrophage polarization.

Alveolar bone regeneration has been strongly linked to macrophage polarization. M1 macrophages aggravate alveolar bone loss, whereas M2 macrophages reverse this process. Berberine (BBR), a natural alkaloid isolated and refined from Chinese medicinal plants, has shown therapeutic effects in treating metabolic disorders. In this study, we first discovered that culture supernatant (CS) collected from BBR-treated human bone marrow mesenchymal stem cells (HBMSCs) ameliorated periodontal alveolar bone loss. CS from the BBR-treated HBMSCs contained bioactive materials that suppressed the M1 polarization and induced the M2 polarization of macrophages in vivo and in vitro. To clarify the underlying mechanism, the bioactive materials were applied to different animal models. We discovered macrophage colony-stimulating factor (M-CSF), which regulates macrophage polarization and promotes bone formation, a key macromolecule in the CS. Injection of pure M-CSF attenuated experimental periodontal alveolar bone loss in rats. Colony-stimulating factor 1 receptor (CSF1R) inhibitor or anti-human M-CSF (M-CSF neutralizing antibody, Nab) abolished the therapeutic effects of the CS of BBR-treated HBMSCs. Moreover, AKT phosphorylation in macrophages was activated by the CS, and the AKT activator reversed the negative effect of the CSF1R inhibitor or Nab. These results suggest that the CS of BBR-treated HBMSCs modulates macrophage polarization via the M-CSF/AKT axis. Further studies also showed that CS of BBR-treated HBMSCs accelerated bone formation and M2 polarization in rat teeth extraction sockets. Overall, our findings established an essential role of BBR-treated HBMSCs CS and this might be the first report to show that the products of BBR-treated HBMSCs have active effects on alveolar bone regeneration.

The Phenotypic and Genotypic Spectrum of CSF1R-Related Disorder in China.

Colony-stimulating factor 1 receptor (CSF1R)-related disorder (CRD) is a rare autosomal dominant disease. The clinical and genetic characteristics of Chinese patients have not been elucidated. The objective of the study is to clarify the core features and influence factors of CRD patients in China. Clinical and genetic-related data of CRD patients in China were collected. Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), and Sundal MRI Severity Score were evaluated. Whole exome sequencing was used to analyze the CSF1R mutation status. Patients were compared between different sexes, mutation types, or mutation locations. A total of 103 patients were included, with a male-to-female ratio of 1:1.51. The average age of onset was (40.75 ± 8.58). Cognitive impairment (85.1%, 86/101) and parkinsonism (76.2%, 77/101) were the main clinical symptoms. The most common imaging feature was bilateral asymmetric white matter changes (100.0%). A total of 66 CSF1R gene mutants (22 novel mutations) were found, and 15 of 92 probands carried c.2381 T > C/p.I794T (16.30%). The MMSE and MoCA scores (17.0 [9.0], 11.90 ± 7.16) of female patients were significantly lower than those of male patients (23.0 [10.0], 16.36 ± 7.89), and the white matter severity score (20.19 ± 8.47) of female patients was significantly higher than that of male patients (16.00 ± 7.62). There is no statistical difference in age of onset between male and female patients. The core manifestations of Chinese CRD patients are progressive cognitive decline, parkinsonism, and bilateral asymmetric white matter changes. Compared to men, women have more severe cognitive impairment and imaging changes. c.2381 T > C/p.I794T is a hotspot mutation in Chinese patients. © 2024 International Parkinson and Movement Disorder Society.

Clinical presentation and diagnosis of adult-onset leukoencephalopathy with axonal spheroids and pigmented glia: a literature analysis of case studies.

Because adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is a rare, rapidly progressive, debilitating, and ultimately fatal neurodegenerative disease, a rapid and accurate diagnosis is critical. This analysis examined the frequency of initial misdiagnosis of ALSP via comprehensive review of peer-reviewed published cases. Data were extracted from a MEDLINE search via PubMed (January 1, 1980, through March 22, 2022) from eligible published case reports/series for patients with an ALSP diagnosis that had been confirmed by testing for the colony-stimulating factor-1 receptor gene (CSF1R) mutation. Patient demographics, clinical symptoms, brain imaging, and initial diagnosis data were summarized descriptively. Categorical data for patient demographics, symptoms, and brain imaging were stratified by initial diagnosis category to test for differences in initial diagnosis based on each variable. Data were extracted from a cohort of 291 patients with ALSP from 93 published case reports and case series. Mean (standard deviation) age of symptom onset was 43.2 (11.6) years. A family history of ALSP was observed in 59.1% of patients. Cognitive impairment (47.1%) and behavioral and psychiatric abnormalities (26.8%) were the most frequently reported initial symptoms. Of 291 total cases, an accurate initial diagnosis of ALSP was made in 72 cases (24.7%) and the most frequent initial misdiagnosis categories were frontotemporal dementia (28 [9.6%]) and multiple sclerosis (21 [7.2%]). Of the 219 cases (75.3%) that were initially mis- or undiagnosed, 206 cases (94.1%) were later confirmed as ALSP by immunohistology, imaging, and/or genetic testing; for the remaining 13 cases, no final diagnosis was reported. Initial diagnosis category varied based on age, family history, geographic region, mode of inheritance, and presenting symptoms of pyramidal or extrapyramidal motor dysfunction, behavioral and psychiatric abnormalities, cognitive impairment, and speech difficulty. Brain imaging abnormalities were common, and initial diagnosis category was significantly associated with white matter hyperintensities, white matter calcifications, and ventricular enlargement. In this literature analysis, ALSP was frequently misdiagnosed. Improving awareness of this condition and distinguishing it from other conditions with overlapping presenting symptoms is important for timely management of a rapidly progressive disease such as ALSP.

Primary Microgliopathy Presenting as Degenerative Dementias: A Case Series of Novel Gene Mutations from India

Introduction: Microglia exert a crucial role in homeostasis of white matter integrity, and several studies highlight the role of microglial dysfunctions in neurodegeneration. Primary microgliopathy is a disorder where the pathogenic abnormality of the microglia causes white matter disorder and leads to a neuropsychiatric disease. Triggering receptor expressed on myeloid cells (TREM2), TYRO protein tyrosine kinase binding protein (TYROBP) and colony-stimulating factor 1 receptor (CSF1R) are genes implicated in primary microgliopathy. The clinical manifestations of primary microgliopathy are myriad ranging from neuropsychiatric syndrome, motor disability, gait dysfunction, ataxia, pure dementia, frontotemporal dementia (FTD), Alzheimer’s dementia (AD), and so on. It becomes imperative to establish the diagnosis of microgliopathy masquerading as degenerative dementia, especially with promising therapies on horizon for the same. We aimed to describe a case series of subjects with dementia harbouring novel genes of primary microgliopathy, along with their clinical, neuropsychological, cognitive profile and radiological patterns. Methods: The prospective study was conducted in a university referral hospital in South India, as a part of an ongoing clinico-genetic research on dementia subjects, and was approved by the Institutional Ethics Committee. All patients underwent detailed assessment including sociodemographic profile, clinical and cognitive assessment, pedigree analysis and comprehensive neurological examination. Subjects consenting for blood sampling underwent genetic testing by whole-exome sequencing (WES). Results: A total of 100 patients with dementia underwent genetic analysis using WES and three pathogenic variants, one each of TREM2, TYROBP, and CSF1R and two variants of uncertain significance in CSF1R were identified as cause of primary microgliopathy. TREM2 and TYROBP presented as frontotemporal syndrome whereas CSF1R presented as frontotemporal syndrome and as AD. Conclusion: WES has widened the spectrum of underlying neuropathology of degenerative dementias, and diagnosing primary microglial dysfunction with emerging therapeutic options is of paramount importance. The cases of primary microgliopathy due to novel mutations in TREM2, TYROBP, and CSF1R with the phenotype of degenerative dementia are being first time reported from Indian cohort. Our study enriches the spectrum of genetic variants implicated in degenerative dementia and provides the basis for exploring complex molecular mechanisms like microglial dysfunction, as underlying cause for neurodegeneration.