G-CSF Research Articles

Real-world use of supportive medications to prevent and manage key adverse events during first-line (1L) treatment of metastatic pancreatic adenocarcinoma (mPDAC) with FOLFIRINOX (FFX), FFX without 5FU bolus, and gemcitabine+nab-paclitaxel (GnP).

695 Background: 1L treatment of mPDAC with FFX or GnP typically requires supportive care to alleviate toxic effects. Two of the most common adverse events are neutropenia, which may be treated with granulocyte colony-stimulating factor (G-CSF), and diarrhea, often treated with atropine and/or loperamide. Medications may be administered as prophylaxis (PPx) or to manage symptoms as they occur. Other methods to improve tolerability include modifying regimens with lower doses or omitted components, such as removing the 5FU bolus from FFX (FFXnb). This study examines PPx vs non-PPx G-CSF, atropine, and loperamide during 1L treatment with FFX, FFXnb, and GnP. Methods: This retrospective observational study utilized Optum Market Clarity claims + EHR linked data. Inclusion criteria were: adult patients (pts) diagnosed with mPDAC between January 1, 2015 and May 31, 2023; initiated treatment (index date) with 1L FFX, FFXnb, or GnP -14 to +90 days from diagnosis; no evidence of other cancers in 6 months pre-index continuous enrollment; at least one EHR record post-index. G-CSF PPx was defined as administration/fill within 7 days of the start of a treatment, inclusive. PPx with atropine or loperamide was defined as administration/fill the day before or day of a treatment; all other use was categorized “non-PPx”. Pts may appear in both PPx and non-PPx categories. Results: Rates of overall use, PPx, and non-PPx for each medication of interest are shown (Table). The overall G-CSF use rate was >3.5 times higher for FFX and FFXnb vs GnP. While ≥70% of FFX/FFXnb pts received PPx, non-PPx still occurred in more than 1 in 8 pts. One-fifth of FFX and FFXnb pts received atropine at any time, vs. just 6% of GnP pts. Atropine PPx was less common than non-PPx for FFX/FFXnb and was rare in GnP pts. Similar to the other medications, loperamide was observed more for FFX and FFXnb vs. GnP, but was the least common of the medications, used by <10% of pts in any regimen. PPx and non-PPx use were similar. Use of the three medications was similar between FFX and FFXnb. Conclusions: In a RW cohort of 1L mPDAC pts, G-CSF and antidiarrheal use was more common in FFX and FFXnb than GnP pts. Most FFX and FFXnb pts received PPx G-CSF, reflecting clinician efforts to manage the high risk of neutropenia with these regimens. Non-PPx G-CSF still occurred in 14–18% of pts. FFXnb had similar rates of concomitant medications as FFX, indicating that removing the 5FU bolus may have minor impact on the need for some supportive medications. Concomitant medication use. Medication 1L FFX(n=511) 1L FFXnb(n=583) 1L GnP(n=958) G-CSF % treated overall 77% 71% 20% % PPx 76% 70% 17% % non-PPx 18% 14% 9% Atropine % treated overall 20% 20% 6% % PPx 11% 9% 2% % non-PPx 15% 16% 5% Loperamide % treated overall 7% 9% 3% % PPx 4% 5% 1% % non-PPx 4% 6% 2%

Impact of Granulocyte Colony-Stimulating Factor (G-CSF) on Clinical Outcomes in Allogeneic Hematopoietic Cell Transplantation: Does Speeding Up Neutrophil Engraftment Make a Difference?

Despite decades of post-allogeneic hematopoietic cell transplantation (HCT) growth factor utilization, its role remains undefined, leading to ongoing debates and research. The theoretical impacts of growth factors have been challenged in numerous studies. In this retrospective cohort study conducted at the Princess Margaret Cancer Centre, we analyzed the clinical outcomes of 509 patients who underwent allogeneic HCT between May 1, 2019, and May 31, 2022. This study aimed to assess the impact of granulocyte colony-stimulating factor (G-CSF) administration posttransplantation on neutrophil and platelet engraftment, incidence of bloodstream infections (BSIs), graft-versus-host disease, engraftment syndrome (ES), and survival metrics including overall survival, nonrelapse mortality, and graft-versus-host disease-free/relapse-free survival. Our findings indicate that G-CSF administration expedited neutrophil engraftment (16 versus 18 d, P = 0.009) and was associated with a decreased incidence of BSI (9.4% versus 31.3%, P = 0.014). However, this benefit was counterbalanced by a significant delay in platelet engraftment (21 versus 17 d, P < 0.001). Multivariate logistic regression analysis identified mismatched donors (odds ratio, 1.72; 95% confidence interval, 1.03-2.88; P = 0.038) and the duration of G-CSF therapy (odds ratio, 1.04; 95% confidence interval, 1.00-1.09; P = 0.038) as independent predictors for the development of ES. Despite these hematological impacts, there was no observed advantage in overall survival, nonrelapse mortality, or graft-versus-host disease-free/relapse-free survival among patients who received G-CSF compared with those who did not. Although G-CSF post-HCT expedited neutrophil engraftment and reduced BSI risk, it did not result in a survival advantage. The association with ES necessitates careful consideration.

The safety and short-term efficacy of induction DCF plus nivolumab therapy in patients with unresectable locally advanced esophageal squamous cell carcinoma.

425 Background: Definitive chemoradiotherapy (dCRT) is the standard treatment for unresectable locally advanced esophageal squamous cell carcinoma (LA-ESCC) based on the results of the JCOG 0303 study. However, the prognosis in this population remains poor with a median overall survival (OS) of 13.1 months, and fistula formation was reported in about 20% of patients who received dCRT. In such a circumstance, a more effective and safer treatment strategy, such as induction chemotherapy followed by conversion surgery or dCRT, was developed. A phase II study of induction chemotherapy consisting of docetaxel (DTX), cisplatin (CDDP), and 5-fluorouracil (5-FU) (DCF) therapy (IC-DCF) showed a conversion rate of 37.5%, promising efficacy with a median OS of 33.8 months and a fistula formation rate of 4.2%. Therefore, we evaluated whether adding nivolumab to IC-DCF (IC-DCF+Nivo) could improve clinical outcomes. Methods: We retrospectively reviewed the medical records of patients with unresectable LA-ESCC who received IC-DCF+Nivo between Nov 2023 and Aug 2024. IC-DCF+Nivo (DTX 70mg/m2 day1, CDDP 70mg/m2 day1, 5-FU 750mg/m2 day1-5, Nivo 360 mg/body day1, every 3 weeks) was administered for 3 courses. We administered prophylactic levofloxacin from day 5 to day 15 during each cycle, and used granulocyte colony-stimulating factor (G-CSF) as a treatment. We evaluated adverse events according to the CTCAE ver5.0 during induction chemotherapy as safety, and objective response rate (ORR) of IC-DCF+Nivo according to the RECIST version 1.1., conversion surgery rate, complete resection (R0) rate, pathological complete response (pCR) rate and clinical CR (cCR) rate after dCRT as efficacy. Results: The median follow-up period was 5.5 months. We identified 23 eligible LA-ESCC patients, median age (range): 65 (44-76), 83% male, PS 0/1: 57%/44%, T status T3/T4b 44%/57%, clinical stage III/IVA/IVB 22%/57%/22%. Seven patients (30%) had supraclavicular lymph node metastases, and the most common unresectable factors were tracheal invasion (52%), bronchial invasion (22%) and aortic invasion (13%) of primary tumor or metastatic lymph nodes. Eighteen patients (78%) completed the 3 courses of IC-DCF+Nivo. The frequent grade 3-4 hematological adverse events were neutropenia (13/23, 57%) and anemia (3/23, 13%), and 1 patient (4.3%) had grade 3 diarrhea as non-hematological toxicity. Febrile neutropenia occurred in 3 patients (13%), and G-CSF was used in 7 patients (30%). One patient (4.3%) experienced primary tumor perforation. The ORR after 3 cycles of induction chemotherapy was 53%. Fourteen patients (61%) achieved resectability, and 10 patients (44%) underwent surgery. R0 resection rate was 90% (9/10), and the pCR rate was 20% (1/10). No one achieved cCR in dCRT. Conclusions: Induction DCF plus nivolumab therapy showed well-tolerated and well conversion rate.

Real-world excess costs associated with hematologic adverse events (hAEs) during first-line (1L) treatment of metastatic pancreatic adenocarcinoma (mPDAC) with FOLFIRINOX (FFX), FFX without 5FU bolus, and gemcitabine+nab-paclitaxel (GnP).

693 Background: During 1L treatment of mPDAC with FFX and GnP, hAEs can harm patients (pts) and accrue excess costs to the health system, i.e., costs beyond those of a treated pt without the hAE. This study examined excess total cost of care (TCoC) and components (total inpatient; outpatient transfusion, granulocyte colony-stimulating factor [G-CSF], and thrombopoietic growth factor [TGF]) for three key hAEs (anemia, neutropenia, thrombocytopenia) during 1L FFX, FFX without 5FU bolus (FFXnb), and GnP. Methods: This retrospective observational study utilized Optum Market Clarity claims + EHR linked data. Inclusion criteria were: adult pts diagnosed with mPDAC between 1/1/2015 and 5/31/2023; initiated 1L FFX, FFXnb, or GnP within -14 to +90 days (index date); ≥6 months pre-index enrollment. hAEs during 1L were detected from lab values and grouped all grades combined. Pts were divided into “hAE groups” and “ref. groups” with and without any of the three hAEs of interest, respectively. Pts were matched 1:1 without replacement based on follow-up time. Costs were measured from the day a lab value indicated an hAE until 30 days later and during the corresponding times for ref. pts, then standardized to per-pt-per-month (PPPM). This study did not adjust for differences in pt characteristics. Results: Means and differences (95% CI) in TCoC following hAE detection for each regimen and hAE of interest are presented (Table). PPPM TCoC was greater for all three regimens and hAEs, driven by 1.6–4.9 times higher inpatient costs. Inpatient costs were $5,614 and $5,665 higher for FFX and FFXnb neutropenia pts vs. ref. groups. Use of outpatient G-CSF at any time during 1L was lower for FFX and FFXnb neutropenia pts, at 76% and 63% vs. 83% and 75% in the ref. groups, respectively. Outpatient TGF and transfusions were rare. Conclusions: All hAEs showed excess PPPM TCoC across regimens, driven by inpatient costs. FFX and FFXnb neutropenia pts had lower overall utilization of outpatient G-CSF but higher inpatient costs following detection of neutropenia, which may represent an opportunity for improved pt management. Excess TCoC of hAEs. AE (any grade) FFX, hAE group(N=199, 127, 157) FFX, ref. group(N=199, 127, 157) Diff. (95% CI) FFXnb, hAE group(N=202, 106, 137) FFXnb, ref. group (N=202, 106, 137) Diff. (95% CI) GnP, hAE group (N=406, 268, 270) GnP, ref. group (N=406, 268, 270) Diff. (95% CI) AnemiaTCoC, mean 30,561 22,310 8,251 (4,396–12,106) 31,866 23,380 8,487(4,007–12,966) 32,662 27,697 4,965(1,320–8,610) NeutropeniaTCoC, mean 30,382 24,952 5,430(-31–10,891) 26,588 22,571 4,017(-1,288–9,322) 32,975 31,051 1,924(-4,629–8,477) ThrombocytopeniaTCoC, mean 30,566 22,535 8,031 (3,895–12,166) 31,434 22,958 8,476(2,962–13,991) 33,863 30,751 3,113(-3,310–9,536)

Serum cytokine profile in women with type 1 diabetes mellitus in the III trimester of pregnancy and in their newborns

Background: Chronic hyperglycemia and abnormal glucose variability are established risk factors for perinatal complications. Abnormal fetal growth and diabetic fetopathy can be associated not only with carbohydrate metabolism disorders, but also with impaired production of cytokines, chemokines, and growth factors that regulate intercellular interactions. Aim: The aim of this study was to evaluate cytokine and growth factor levels in the blood serum of pregnant women with type 1 diabetes mellitus and in umbilical cord blood of their neonates, as well as assessing the associations of these levels with the development of fetal macrosomia and diabetic fetopathy. Materials and methods: This prospective study included 88 women with a singleton pregnancy and cesarean delivery. All the patients provided informed consent for participation. The study groups comprised individuals with type 1 diabetes mellitus (n = 32) and non-impaired glucose tolerance (n = 56). The levels of interferons alfa 2 and gamma, monokine induced by interferon-gamma, interleukin-1α, -1β, -2, -3, -4, -5, -6, -7, -8, -9, -10, -13, -15, -16, -17, -18, interleukin-1 receptor antagonist, interleukin-2 receptor alpha subunits, p40 and p70 subunits of interleukin-12, monocyte chemotactic protein-1, -3, macrophage inflammatory protein-1α, -1β, leukemia inhibitory factor, macrophage ingibitory factor, nerve growth factor beta, stem cell factor, cutaneous T-cell-attracting chemokine, growth-regulated oncogene alpha, hepatocyte growth factor, stem cell growth factor beta, stromal cell-derived factor-1 alfa, interferon gamma inducible protein 10, platelet-derived growth factor-bb, vascular endothelial growth factor, basic fibroblast growth factor, eotaxin, chemokine ligand 5, and macrophage, granulocyte, and granulocyte/macrophage colony-stimulating factors, tumor necrosis factor alfa and beta, and tumor necrosis factor-related apoptosis-inducing ligand were measured with multiplex immunoassay in the blood serum of women at 36–40 weeks of gestation and in the umbilical cord blood serum of their neonates. Results: Cytokine levels varied widely in the both study groups. Women with type 1 diabetes mellitus had higher levels of interleukin-1β, -6, -17, basic fibroblast growth factor, macrophage inflammatory protein-1α, -1β, and lower levels of interleukin-1ra compared to the control group. Their neonates had higher levels of interleukin-6, -8, granulocyte colony-stimulating factor, macrophage inflammatory protein-1α, -1β. Higher levels of interleukin-1β, -6, -17, and basic fibroblast growth factor in the blood serum of the women were associated with time in range less than 70% and the development of diabetic fetopathy. Time in range was inversely associated with the interferon gamma / interleukin-5 (ρ = −0.531; p = 0.013), interferon gamma / interleukin-10 (ρ = −0.441; p = 0.045), interleukin-2 / -10 (ρ = −0.473; p = 0.030), interleukin-1β / -10 (ρ = −0.561; p = 0.008), interleukin-1β / -1ra (ρ = −0.635; p = 0.002), and interleukin-6 / -10 (ρ = −0.540; p = 0.012) ratios. Conclusions: Cytokine profile alterations were the most pronounced in pregnant women with non-target time in range values and in those with diabetic fetopathy development. Serum cytokine levels correlate with glycemic profile parameters but are considered non-specific markers of pregnancy complications.

P-1013. Incidence and Risk Factors for Fungal Infection after CD19-targeted Chimeric Antigen Receptor T cell Therapy for Non-Hodgkin Lymphoma

Abstract Background Chimeric antigen receptor T cell therapy (CAR T) is highly effective for the treatment of non-Hodgkin lymphoma (NHL); however, recipients are at risk of infectious complications. Infections following CAR T are predominantly bacterial or viral; nevertheless, a subset of recipients develop invasive fungal disease (IFD). The limited evidence guiding antifungal prophylaxis (PPx) led us to investigate the incidence and risk factors for IFD at our center. Legend: DLBCL=diffuse large B-cell lymphoma; MCL=mantle cell lymphoma; tFL=transformed follicular lymphoma; PMBCL=primary mediastinal large B-cell lymphoma; HGBCL= high-grade B-cell lymphoma; autoHCT=autologous hematopoietic cell transplant; alloHCT=allogeneic hematopoietic cell transplant; CAR =chimeric antigen receptor; Axi-cel=axicabtagene ciloleucel; Brexu-cel=brexucabtagene autoleucel; Tisa-cel=tisagenlecleucel; CRS=cytokine release syndrome; ICANS= immune effector cell-associated neurotoxicity syndrome; IEC-HS/MAS= immune effector cell-associated HLH-like syndrome/macrophage activation syndrome; IgG=immunoglobulin G; G-CSF=granulocyte colony-stimulating factor Methods Adults with NHL who received commercial CAR T between January 2018 and February 2021 at MD Anderson Cancer Center were retrospectively identified. Demographics, oncologic history, and complications related to therapy were collected. Consensus definitions were used to define probable and proven IFD. Patients were observed for 1 year after CAR T or until death, whichever occurred first. Routine antifungal PPx was administered per institutional protocol. This database was built with funds from Merck. Legend: IFD=invasive fungal disease; PJP=Pneumocystis jirovecii pneumonia Results A total of 219 patients were identified (Table 1). Most patients had an underlying diagnosis of diffuse large B cell lymphoma (73.5%) and received axicabtagene ciloleucel (93.6%). Therapy related toxicities included: cytokine release syndrome (CRS) (88.1%), immune effector cell-associated neurotoxicity syndrome (ICANS) (58.9%) and immune effector cell-associated hemophagocytic lymphohistiocytosis-like syndrome (IEC-HS) (8.2%). Sixteen patients (7.3%) developed IFD, including 9 (4.1%) mold infections (Table 2). In univariate analysis (Table 3a), IEC-HS, grade 4 CRS, grade 4 ICANS, and any infection within 30 days prior to CAR T were associated with IFD. In multivariate analysis (Table 3b), grade 4 CRS, grade 4 ICANS, and any infection within 30 days prior to CAR T were independent predictors for IFD. Factors associated with mold infections in univariate analysis (Table 3c) included IEC-HS, grade 4 CRS, grade 4 ICANS, steroid use, prior hematopoietic cell transplantation (HCT), and any infection documented within 30 days prior to CAR T. Legend: IFD=invasive fungal disease; cHR= Crude hazard ratio; 95% CI= 95% Confidence interval; aHR= Adjusted hazard ratio; CAR T=Chimeric antigen receptor T cell therapy; HCT=autologous hematopoietic cell transplant; CRS=cytokine release syndrome; ICANS= immune effector cell-associated neurotoxicity syndrome; IEC-HS/MAS= immune effector cell-associated HLH-like syndrome/macrophage activation syndrome; ANC=absolute neutrophil count; ALC=absolute lymphocyte count. Note:* indicates the variable was treated as a time-dependent variable in the analysis. Multivariate analysis on mold infection was not performed due to the small number of the events. Conclusion IFD after CAR T for NHL is uncommon. Higher grade toxicities and their treatment, as well as prior HCT and prior infections, may identify those at highest risk. Additional studies are needed to individualize antifungal PPx strategies in this population. Disclosures Charles Gaulin, MBBS, ADC Therapeutics: Honoraria|DeciBio: Advisor/Consultant|Sanofi: Honoraria Roy F. Chemaly, MD/MPH, AiCuris: Advisor/Consultant|AiCuris: Grant/Research Support|Ansun Pharmaceuticals: Advisor/Consultant|Ansun Pharmaceuticals: Grant/Research Support|Astellas: Advisor/Consultant|Eurofins-Viracor: Grant/Research Support|InflaRX: Advisor/Consultant|Janssen: Advisor/Consultant|Karius: Advisor/Consultant|Karius: Grant/Research Support|Merck/MSD: Advisor/Consultant|Merck/MSD: Grant/Research Support|Moderna: Advisor/Consultant|Oxford Immunotec: Advisor/Consultant|Oxford Immunotec: Grant/Research Support|Roche/Genentech: Advisor/Consultant|Roche/Genentech: Grant/Research Support|Shinogi: Advisor/Consultant|Takeda: Advisor/Consultant|Takeda: Grant/Research Support|Tether: Advisor/Consultant Fareed Khawaja, MBBS, Eurofins Viracor: Grant/Research Support|Symbio: Grant/Research Support

To Study Level of Gonadal Hormone in Cancer Patients Receiving Chemotherapy

Objectives: The objective of this study was to study the alteration in levels of testosterone, luteinising hormone (LH), follicle-stimulating hormone (FSH) and prolactin in male cancer patients’ post-chemotherapy who have been started on drugs causing grade III or more haematological toxicity requiring granulocyte colony-stimulating factor support. Materials and Methods: An open-labelled prospective single-centre study for the assessment of Gonadal dysfunction in male cancer patients on chemotherapy from August 2010 to October 2012. Results: Patients had a significant alteration in testosterone (23% n = 11), LH (52.08% n = 25), FSH (83.3% n = 40) and prolactin levels (22.92% n = 11) post-chemotherapy, which was statistically significant (P &lt; 0.05). The high dose, as well as the low dose Cisplatin group, were associated with substantial gonadal dysfunction in testicular tumour patients. Hence, regimens using low-dose cisplatin at the cost of compromising the cure rate for the motive of preservation of fertility are not warranted. There was no statistically significant difference found in alteration in mean testosterone, mean LH or mean FSH Prolactin levels among cisplatin, cyclophosphamide or combined cisplatin and cyclophosphamide chemotherapy group, at least in the initial weeks of post-chemotherapy. Conclusion: There is a significant gonadal dysfunction in cancer patients receiving cytotoxic chemotherapy. Hence, increasing awareness to prevent, detect and treat expected long-term toxicities is crucial. Prechemotherapy cryopreservation of semen and assisted reproductive technologies are good options to describe the rate of involuntary infertility without affecting the survival of patients by decreasing doses of chemotherapy.

Healthcare Resource Utilization Associated with Leukopenia and Neutropenia in Kidney Transplant Recipients Receiving Valganciclovir in the United States

Background: Cytomegalovirus prophylaxis in kidney transplant recipients (KTRs) is limited by post-transplant neutropenia and leukopenia (PTN/PTL). Despite its clinical significance, the healthcare resource utilization (HCRU) related to PTN/PTL remains poorly characterized. Objective: To evaluate HCRU among KTRs taking valganciclovir during their first year post-transplant. Methods: Using TriNetX Dataworks-USA, a federated, de-identified electronic medical record database, we identified adult KTRs who underwent their first kidney transplant from January 2012 to September 2020. All eligible patients were followed for 1 year. PTN/PTL was defined as absolute neutrophil count less than 1000/μL or white blood cell count less than 3500/μL. Multivariable logistic/Poisson regression models were used to assess the association between PTN/PTL and various HCRU types. Results: A total of 8791 KTRs were identified, of whom 6219 (70.7%) developed PTN/PTL at a mean of 5.7 months post-transplantation. Hospitalizations, rehospitalizations, emergency room visits, outpatient appointments, packed red blood cell transfusions, and granulocyte-colony stimulating factor administration were more prevalent among KTRs with PTN/PTL (61.1% vs 49.5%, 24.5% vs 14.1%, 35.2% vs 28.9%, 30.4 vs 26.2 visits, 22.3% vs 17.6%, 23.4% vs 2.2%, respectively; P &lt; .001). Adjusted analyses confirmed that PTN/PTL correlated with increased HCRU across all categories. Conclusions: KTRs who developed PTN/PTL had significantly higher HCRU. Further studies are needed to evaluate strategies addressing PTN/PTL for KTRs.

Deciphering the therapeutic effects of Xiyanping injection: insights into pulmonary and gut microbiome modulation, SerpinB2/PAI-2 targeting, and alleviation of influenza a virus-induced lung injury

Infection with Influenza A virus (IAV) induces severe inflammatory responses and lung injury, contributing significantly to mortality and morbidity rates. Alterations in the microbial composition of the lungs and intestinal tract resulting from infection could influence disease progression and treatment outcomes. Xiyanping (XYP) injection has demonstrated efficacy in clinical treatment across various viral infections. However, its specific effects and mechanisms against IAV remain unclear. In this study, we established an IAV infection mice model, and utilized 16 S rRNA sequencing, RNA sequencing, protein chips, and molecular docking, to investigate the mechanisms of XYP injection on altering pulmonary and gut microbiota, and identifying its target sites. We revealed that XYP injection significantly reduced mortality, weight loss, lung viral titers, and lung pathology in IAV-infected mice. XYP injection down-regulated the activity of malondialdehyde, and the levels of interleukin (IL)-1β, IL-5, IL-6, tumor necrosis factor-α, IL-18, IL-15, granulocyte colony-stimulating factor, IL-9, chemokine (C-C motif) ligand-5, and C-X-C motif chemokine ligand 5, while up-regulated the activities of glutathione peroxidase reactive and superoxide dismutase, and the level of interferon-γ. The diversity of the pulmonary and gut microbiota was altered slightly after XYP injection. The linear discriminant analysis of the gut microbes revealed a higher proportion of potentially beneficial bacteria, including Akkermansia, Parabacteroides goldsteinii, Defluviitaleaceae, Oscillospirales, and Eubacterium_coprostanoligenes_group characterized the XYP group. Peritoneal macrophage RNA sequencing highlighted Serpinb2 as the most significantly regulated gene by XYP injection, along with consistent changes in multiple downstream Th2 structure genes. KEGG pathway analysis indicated significant modifications in genes associated with influenza A, mitogen-activated protein kinase signaling, nuclear factor kappa-B signaling, and apoptosis following XYP injection. Finally, human protein chips and molecular docking were carried out to confirm the binding of the main component of XYP injection, andrographolide, with SERPINB2/PAI-2 protein. Overall, our study provides valuable insights into the therapeutic potential of XYP injection in treating influenza, highlighting its multifaceted effects on host microbiota and immune responses, and pinpointing SerpinB2/PAI-2 as the target for XYP injection in exerting anti-inflammatory and antiviral therapeutic mechanisms.

Sorting Out the SOCS Genes and Their Role in Macrophage Activation.

The suppressors of cytokine signaling (SOCS) genes were first described in a group of articles published in 1997. Since that time, much has been learned about the functional activities mediated by the corresponding proteins encoded by the SOCS genes. The SOCS gene family contains eight members: SOCS1 through SOCS7 and a highly related gene known as CISH (cytokine-inducible SH2-containing protein). Although much is known about the ability of the SOCS proteins to autoregulate responses to individual cytokines, much less is known about the ability of the SOCS proteins to cross-regulate cytokine signaling. The studies described in a new report by Bidgood et al. in this issue of JICR demonstrate that SOCS1 expression induced by one cytokine, interferon (IFN)-γ, can cross-regulate signaling induced by another cytokine, granulocyte macrophage colony-stimulating factor (GM-CSF), in murine bone marrow-derived macrophages. The authors show that the ability of SOCS1 to inhibit cytokine signaling is dose- and time-dependent. SOCS1 must reach a critical threshold level before it can exert a marked inhibitory effect on autocrine signaling through the IFN-γ receptor or paracrine signaling through the GM-CSF receptor.

The impact of cytokines and tumour-conditioned medium on the properties of murine invitro generated myeloid-derived suppressor cells.

Myeloid-derived suppressor cells (MDSCs) are a heterogeneous group of immature myeloid cells playing a critical role in immune suppression. Invitro-generated MDSCs are a convenient tool to study the properties of tumour-associated MDSCs. Here, we compared six protocols for invitro generation of functional mouse MDSCs from bone marrow progenitors. The protocols included granulocyte-macrophage colony-stimulating factor (GM-CSF) alone or in combination with interleukin-6 (IL-6) or granulocyte colony-stimulating factor (G-CSF), with or without a tumour-conditioned medium (TCM) derived from B16-F10 melanoma. Obtained MDSCs were characterized by morphology, phenotype, gene expression of key immunosuppressive factors, and invitro suppression of T cell proliferation. All tested protocols yielded approximately 25% monocytic and 50% polymorphonuclear MDSCs. Protocols using IL-6 generated MDSCs with reduced maturation and differentiation status, upregulated Arg1 and Nos1 mRNA expression, increased levels of Arg-1 and TGF-β proteins and enhanced ROS production compared to the other protocols. All tested protocols yielded MDSCs that efficiently inhibited T cell proliferation invitro, with some advantage for the GM-CSF and G-CSF + GM-CSF protocols. Interestingly, a combination of protocols with B16-F10-derived TCM resulted in the generation of MDSCs with reduced immunosuppressive properties. Our results provide valuable insights into the optimal conditions for invitro generation of MDSCs with specific immunosuppressive properties.

Silibinin, a PLC-β3 inhibitor, inhibits mast cell activation and alleviates OVA-induced asthma.

The immunoglobulin E (IgE) receptor FcεRI (Fc epsilon RI) plays a crucial role in allergic reactions. Recent studies have indicated that the interaction between FcεRIβ and the downstream protein phospholipase C beta 3 (PLCβ3) leads to the production of inflammatory cytokines. The aim of this study was to develop small molecules that inhibit the protein-protein interactions between FcεRIβ and PLCβ3 to treat allergic inflammation. Additionally, PLCβ3 has emerged as a potential target protein for treating allergic inflammation. In this study, we employed a virtual screening technique to search the Taiwan Traditional Chinese Medicine Database, followed by a second screening using absorption, distribution, metabolism, excretion, and toxicity (ADMET). Among the compounds screened, silibinin exhibited the best performance, forming strong hydrogen bond interactions with residues of PLCβ3, with a binding free energy of -119.277 kcal/mol. Therefore, silibinin effectively blocked the interaction between FcεRIβ and PLCβ3. Silibinin reduced the production of allergic inflammatory cytokines, including cytokine-induced neutrophil chemoattractant 2a (CINC-2a), interleukin-2 (IL-2), cytokine-induced neutrophil chemoattractant 1 (CINC-1), interleukin 1α (IL-1α), macrophage inflammatory protein 3 alpha (MIP3α), interferon γ (IFN-γ), activin A, granulocyte macrophage colony stimulating factor (GM-CSF), intercellular adhesion molecule-1 (ICAM-1), interleukin 4 (IL-4), interleukin 13 (IL-13), Fas ligand (FasL) and tumor necrosis factor alpha (TNF-α), without inducing cytotoxicity. Furthermore, in studies of IgE-mediated allergic responses, silibinin also decreased the expression of surface IgE receptors (FcεRIs). Moreover, silibinin effectively alleviated allergen-induced asthma responses and reduced the infiltration of inflammatory immune cells into the lungs of an OVA-induced allergic airway inflammation mouse model. Taken together, these results demonstrate the potential antiallergic mechanism of silibinin both in vitro and in vivo, making it a promising candidate for the development of asthma therapeutics.

Single-cell transcriptomics predict novel potential regulators of acute epithelial restitution in the ischemia-injured intestine.

Intestinal ischemic injury damages the epithelial barrier predisposes patients to life-threatening sepsis unless that barrier is rapidly restored. There is an age-dependency of intestinal recovery in that neonates are the most susceptible to succumb to disease of the intestinal barrier versus older patients. We have developed a pig model that demonstrates age-dependent failure of intestinal barrier restitution in neonatal pigs which can be rescued by the direct application of juvenile pig mucosal tissue, but the mechanisms of rescue remain undefined. We hypothesized that by identifying a subpopulation of restituting enterocytes by their expression of cell migration transcriptional pathways, we can then predict novel upstream regulators of age-dependent restitution response programs. Superficial mucosal epithelial cells from recovering ischemic jejunum of juvenile pigs underwent single cell transcriptomics and predicted upstream regulator CSF-1 was interrogated in our model. A subcluster of absorptive enterocytes expressed several cell migration pathways key to restitution. Differentially expressed genes in this subcluster predicted their upstream regulation by colony stimulating factor-1 (CSF-1). We validated age-dependent induction of CSF-1 by ischemia and documented that CSF-1 and CSF1R co-localized in ischemic juvenile, but not neonatal, wound-adjacent epithelial cells and in the restituted epithelium of juveniles and rescued neonates. Further, the CSF-1 blockade reduced restitution in vitro, and CSF-1 improved barrier function in injured neonatal pig in preliminary ex vivo experiments. These studies validate an approach to inform potential novel therapeutic targets, such as CSF-1, to improve outcomes in neonates with intestinal injury in a unique pig model.

Amelioration of liver fibrosis with autologous macrophages induced by IL-34-based condition

BackgroundFor the treatment of liver fibrosis, several novel cell therapies have been proposed. Autologous macrophage therapy has been reported as one of the promising treatments. So far, most studies have used colony-stimulating factor 1 (CSF-1) to induce the differentiation of macrophage progenitor cells. The receptor for CSF-1, CSF-1R possesses another ligand, interleukin 34. However, the therapeutic capacity for liver fibrosis by interleukin 34-induced macrophages has not been evaluated.MethodsWe have employed acute (bile duct ligation) and chronic (administration of carbon tetrachloride or thioacetamide) liver fibrosis models. Using these models, we evaluated the therapeutic capacity of macrophages induced by interleukin 34-based conditions. In most experiments, interleukin 4 was also added to the differentiation process to induce alternative-activated macrophages. As a mechanism analysis, we have examined liver inflammation and damage, the status of stellate cells, and the immunosuppressive capacity of the macrophages. Human macrophages were differentiated from CD14+ monocytes and analyzed.ResultsIn both acute and chronic liver damage experiments, interleukin 34-induced macrophages significantly ameliorated liver fibrosis. The addition of interleukin 4 to the differentiation process resulted in an increase of obtained macrophages and a bias to alternative activated macrophages (so-called M2). The alternative activated macrophages (M2-type) showed a reproducible therapeutic effect of liver fibrosis with a suppression of parameters of liver inflammation and damage, stellate cells, and T cell activation. Similar macrophages could be differentiated from human CD14+ monocytes in the presence of interleukin 34 plus interleukin 4, and a therapeutic effect was observed using a humanized mouse model.ConclusionsInterleukin 34-induced macrophages, particularly when additionally stimulated with interleukin 4, significantly ameliorated the liver fibrosis.

Decreased PD-L1 contributes to preeclampsia by suppressing GM-CSF via the JAK2/STAT5 signal pathway

Programmed cell death protein 1 (PD-1) and its ligand PD-L1 have been detected at the materno-embryonic interface in both human and murine pregnancy models. However, research regarding the PD-1/PD-L1 signal in preeclampsia (PE) is limited. In the present investigation, 30 normal pregnant females and 30 PE patients were enrolled. Cellular functional experiments were performed in two trophoblast cell lines by transfection with lentiviral vectors for overexpression and down-regulation of PD-L1. The placental expressions of PD-1, PD-L1, and granulocyte macrophage colony-stimulating factor (GM-CSF) exhibited a notable reduction in PE cases compared with healthy pregnancies. Cellular functional experiments indicated that excessive PD-L1 expression significantly enhanced trophoblast migratory, invasive, and proliferative capabilities while inhibiting cell apoptosis. Additionally, the administration of lentivirus-mediated PD-L1 overexpression could alleviate clinical symptoms (hypertension, proteinuria) of PE-like rats. Therefore, decreased PD-L1 may contribute to PE by inhibiting GM-CSF via activating the JAK2/STAT5 pathway. Our study provides a novel pathway that can be targeted for the therapy of this disease.

P0252 Mild Crohn’s disease is associated with altered sphingolipid metabolism and reduced neutrophilic inflammation

Abstract Background Around 20-30% of patients with Crohn's disease (CD) experience a mild disease course. However, identifying patients likely to maintain a mild course is challenging. This study aimed to identify molecular features of mild CD by exploring biological datasets from two large patient cohorts. Methods Patients with mild CD were analysed from two longitudinal cohorts: the Mount Sinai Crohn's and Colitis Registry (MSSCR) observational cohort and the Ocean State Crohn’s and Colitis Registry (OSCCAR) inception cohort. Baseline assessments in both cohorts included biological/immunomodulator use, history of IBD-related surgeries, and complications (stricturing, penetrating, and perianal CD). Mild CD was defined by the absence of these variables at baseline and after five years of follow-up (Fig. 1A). The remaining patients were mild at baseline but progressed during follow-up. Patients with mild CD and those who progressed after being mild at baseline were evaluated for potential molecular biomarkers including circulating metabolites (metabolomics), immune cell profiles (multiparametric flow cytometry), serum proteomics, and intestinal transcriptomics. Results In total, 100 (MSSCR) and 152 (OSCCAR) patients were characterized, with 55 (55.0%) and 74 (48.7%) classified as mild CD, respectively. Intestinal transcriptomic analysis revealed downregulation of genes associated with neutrophilic inflammation and innate immunity, including SAA, DUOXA2, FCGR3B, LCN2, S100A8, and AQP9 (Fig. 1B). Flow cytometry showed lower frequencies of neutrophils (CD16+), lower neutrophil-to-lymphocyte ratios (NLR), and higher counts of NK and total B cells in mild CD (Fig. 1C). Metabolomic analysis identified ceramide- and sphingomyelin compounds (particularly acyl-CoA chain lengths C14-16 and C22-26) as positively associated with mild CD (odds ratios [OR] &amp;gt;2.0), whereas primary and secondary bile acids and amino acid derivatives from lysine, tyrosine, and proline metabolism were inversely associated (Fig. 1D). Proteomic analysis in OSCCAR demonstrated associations between C-X-C motif chemokine ligand 9 (CXCL9), macrophage colony stimulating factor (CSF-1), and TGF-β1 latency-associated peptide (LAP-TGF-β), and an inverse association for Delta and Notch-like epidermal growth factor-related receptor (DNER) with mild CD (adjusted P&amp;lt;0.05) (Fig. 1E). Conclusion This study provides multi-omics-based biomarker signatures of mild CD, which primarily relate to altered ceramide- and sphingomyelin metabolites and lower systemic neutrophilic inflammation. Future studies focusing on the biological significance and potential clinical impact on disease monitoring and therapeutic strategies are warranted.

Hidradenitis suppurativa.

Hidradenitis suppurativa is a chronic inflammatory disease characterised by painful, deep-seated nodules, abscesses, and draining tunnels in the skin of axillary, inguinal, genitoanal, or inframammary areas. In recent years, the body of knowledge in hidradenitis suppurativa has advanced greatly. This disorder typically starts in the second or third decade of life. The average worldwide prevalence is 1% but varies geographically. Hidradenitis suppurativa has a profound negative effect on patients' quality of life and on the gross value added to society. Comorbidities (eg, metabolic syndrome, inflammatory arthritis, and inflammatory bowel disease) frequently accompany skin alterations, because of systemic inflammation. Pathogenesis of hidradenitis suppurativa is complex and includes innate immune mechanisms (eg, macrophages, neutrophils, IL-1β, tumour necrosis factor [TNF], and granulocyte colony-stimulating factor), T-cell mechanisms (eg, IL-17 and IFN-γ), and B-cell mechanisms (eg, associated with dermal tertiary lymphatic structures and autoantibodies). Chronic inflammation leads to irreversible skin damage with tunnel formation and morbid scarring. Current treatment includes drug therapy (for the initial, purely inflammatory phase), combined drug and surgical therapy (for the destructive phase), or surgery alone (for the burnout phase). The first systemic therapies approved for hidradenitis suppurativa targeting TNF (adalimumab) and IL-17 (secukinumab and bimekizumab) have expanded drug therapy options for moderate-to-severe disease, which were previously mainly restricted to oral antibiotics. Moreover, there is a robust pipeline of immunomodulatory drugs in various stages of development for hidradenitis suppurativa. Aims of management should include early intervention to prevent irreversible skin damage, adequate control of symptoms including pain, and mitigation of extra-cutaneous comorbidities, all requiring early diagnosis and an interdisciplinary, holistic and personalised approach.

P0558 An Integrative Blood-Based Risk Score Predicts Development of Crohn's Disease

Abstract Background Individual blood markers have been associated with future risk of Crohn's disease (CD). However, there is a need to understand which combination of biomarkers will be most predictive to facilitate CD prevention trial recruitment. We aimed to develop and validate a blood-based integrative risk score for CD development. Methods We analyzed data from 200 patients with CD and 100 age, sex, and race-matched healthy controls (HC) from PREDICTS, a nested case-control study of US active-component service members (ACSM). Longitudinal serum samples were collected at four time points up to 6+ years prior to diagnosis. Anti-microbial antibodies (Prometheus), proteomic markers (Olink inflammation panel), and anti-granulocyte macrophage-colony stimulating factor autoantibodies (anti-GM-CSF) were assayed in each sample. Participants were randomly divided into equal sized training and testing sets. Time-varying trajectories of marker abundance were estimated for each biomarker. Logistic regression modeled disease status as a function of each marker for different time points and multivariate modeling was performed via logistic lasso regression. A risk score to predict CD onset within 2 years was developed using penalized mixed-effects logistic regression. Prediction models were fit on the testing set and predictive performance evaluated via receiver operating characteristic (ROC) curves and area under the curve (AUC). Results The AUCs of individual markers were dynamic over time, with some having stable predictive capacity and others increasing or decreasing closer to time of diagnosis (Figure 1A). The AUCs for a model combining antibodies (Prometheus and anti-GM-CSF) and proteins were consistently above 0.8 starting 4 years prior to diagnosis, primarily driven by proteomic markers (Figure 1B). An integrative model predicting CD onset within 2 years incorporated 10 biomarkers and significantly associated with CD onset (Figure 1C). The model resulted in an AUC of 0.87 with a specificity of 99% and positive predictive value of 84% at a classification threshold of 0.7 (Figure 1D). Stratifying the integrative model scores into quartiles revealed a clear gradient in CD incidence within 2 years, increasing from 2% in the first quartile to 57.7% in the fourth quartile (Figure 1E). The relative risk (RR) for developing CD among ACSM in the top quartile, compared to the lower quartiles, was 10.4 (95% CI 6.9, 15.6). Conclusion An integrative blood-based model combining anti-microbial antibodies and proteins predicts onset of CD. Serologic and proteomic markers have dynamic changes years before diagnosis, highlighting the varying preclinical phases of CD and how leveraging biomarker dynamics can predict time to diagnosis. Figure 1

Simple dysmood disorder, a mild subtype of major depression, is not an inflammatory condition: Depletion of the compensatory immunoregulatory system.

A recent study conducted by the laboratory of the first author revealed that major depression is composed of two distinct subtypes: major dysmood disorder (MDMD) and simple dysmood disorder (SDMD). The latter is a less severe phenotype with fewer aberrant biological pathways. MDMD, but not SDMD, patients were identified to have highly sensitized cytokine/growth factor networks using stimulated whole blood cultures. However, no information regarding serum cytokines/chemokines/growth factors in SDMD is available. This case-control study compares 48 serum cytokines/chemokines/growth factors in academic students with SDMD (n=64) and first episode (FE)-SDMD (n=47) to those of control students (n=44) using a multiplex assay. Both FE-SDMD and SDMD exhibited a notable inhibition of immune profiles, such as the compensatory immunoregulatory response system (CIRS) and alternative M2 macrophage and T helper-2 (Th-2) profiles. We observed a substantial reduction in the serum concentrations of five proteins: interleukin (IL)-4, IL-10, soluble IL-2 receptor (sIL-2R), IL-12p40, and macrophage colony-stimulating factor. A considerable proportion of the variability observed in suicidal behaviors (26.7%) can be accounted for by serum IL-4, IL-10, and sIL-2R (all decreased), CCL11 (eotaxin) and granulocyte CSF (both increased). The same biomarkers (except for IL-10), accounted for 25.5% of the variance in SDMS severity. A significant correlation exists between decreased levels of IL-4 and elevated ratings of the brooding type of rumination. SDMD is characterized by the suppression of the CIRS profile, which signifies a disruption of immune homeostasis and tolerance, rather than the presence of an inflammatory response.

Label-free Detection and Characterization of Metabolism in Fresh and Cryopreserved Macrophages.

Cryopreservation is a widely used technique to preserve biological samples for extended periods of time at low temperatures. Even though it is known to have significant effects on cell viability, its effect on their metabolism remains unexplored. Studying how cryopreservation influences the metabolism of cells is important to guarantee the reliability of samples transported between sites for analysis. Optical metabolic imaging allows for the study of cellular metabolism in a label-free manner by using the autofluorescence properties of nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD), two metabolic coenzymes. The goal of this research is to study the metabolic changes in macrophages after cryopreservation and compare these results with freshly isolated macrophage samples to evaluate if the metabolic data is retained after cryopreservation. The metabolism of macrophages was analyzed with fluorescence lifetime imaging microscopy (FLIM) using a multiphoton microscope. Monocytes were isolated from human whole blood and separated into two groups. In Group 1, freshly isolated monocytes were differentiated into macrophages using macrophage-colony stimulating factor (M-CSF) over 8 days. In Group 2, isolated monocytes were cryopreserved after separation from whole blood and then thawed and stimulated with M-CSF for 8 days. Single-cell analysis showed there are significant changes in FLIM parameters between the groups suggesting that the metabolic data of macrophages is altered after a cryopreservation and cell thawing cycle. Our research bridges the current gap by studying the metabolic changes of cells after cryopreservation using a non-invasive and label-free imaging technique.