Lina Kozhaya

Objective: Chronic rhinosinusitis (CRS) impacts an estimated 5% to 15% of people worldwide, incurring significant economic healthcare burden. There is a urgent need for the discovery of predictive biomarkers to improve treatment strategies and outcomes for CRS patients. Study design: Cohort study of CRS patients and healthy controls using blood samples. Setting: Out-patient clinics. Methods: Whole blood samples were collected for flow cytometric analysis. Mechanistic studies involved the transfection of human primary T cells and Jurkat cells. Results: Our analysis began with a 63-69 year-old female patient diagnosed with refractory CRS,. Despite undergoing multiple surgeries, she continually faced sinus infections. Whole exome sequencing pinpointed a heterozygous IL-12Rb1 mutation situated in the linker region adjacent to the cytokine binding domain. When subjected to IL-12 stimulation, the patient’s CD4 T-cells exhibited diminished STAT4 phosphorylation. However, computer modeling or T-cell lines harboring the same IL-12 receptor mutation did not corroborate the hypothesis that IL-12Rb could be responsible for the reduced phosphorylation of STAT4 by IL-12 stimulation. Upon expanding our investigation to a broader CRS patient group using the pSTAT4 assay, we discerned a subset of refractory CRS patients with abnormally low STAT4 phosphorylation. The deficiency showed improvement both in-vitro and in-vivo after exposure to Latilactobacillus sakei (aka Lactobacillus sakei ), an effect at least partially dependent on IL-12. Conclusion: In refractory CRS patients, an identified STAT4 defect correlates with poor clinical outcomes after sinus surgery, which can be therapeutically targeted by Latilactobacillus sakei treatment. Prospective double-blind placebo-controlled trials are needed to validate our findings.

XLS file, 17K, Patient demographics, tumor characteristics, treatment, occurrence and severity of local and systemic adverse events and anti-tumor response per patient (n=10).

PDF file, 282K, A (non-responder 1): In situ TILs analysis by IHC in a patient with minimal T cell infiltrate before treatment (H&E stain and IHC for CD3, CD4, CD8 and FoxP3, 200x). B: (non-responder 2): In situ TILs analysis by IHC in a patient with moderate T cell infiltrate before imiquimod treatment. (H&E stain and IHC for CD3, CD4, CD8 and FoxP3, 200x).

Purpose: Skin metastases of breast cancer remain a therapeutic challenge. Toll-like receptor 7 agonist imiquimod is an immune response modifier and can induce immune-mediated rejection of primary skin malignancies when topically applied. Here we tested the hypothesis that topical imiquimod stimulates local antitumor immunity and induces the regression of breast cancer skin metastases.Experimental Design: A prospective clinical trial was designed to evaluate the local tumor response rate of breast cancer skin metastases treated with topical imiquimod, applied 5 d/wk for 8 weeks. Safety and immunologic correlates were secondary objectives.Results: Ten patients were enrolled and completed the study. Imiquimod treatment was well tolerated, with only grade 1 to 2 transient local and systemic side effects consistent with imiquimod's immunomodulatory effects. Two patients achieved a partial response [20%; 95% confidence interval (CI), 3%–56%]. Responders showed histologic tumor regression with evidence of an immune-mediated response, showed by changes in the tumor lymphocytic infiltrate and locally produced cytokines.Conclusion: Topical imiquimod is a beneficial treatment modality for breast cancer metastatic to skin/chest wall and is well tolerated. Importantly, imiquimod can promote a proimmunogenic tumor microenvironment in breast cancer. Preclinical data generated by our group suggest superior results with a combination of imiquimod and ionizing radiation and we are currently testing in patients whether the combination can further improve antitumor immune and clinical responses. Clin Cancer Res; 18(24); 6748–57. ©2012 AACR.

Objectives: Despite advances in antibody treatments and vaccines, COVID‐19 caused by SARS‐CoV‐2 infection remains a major health problem resulting in excessive morbidity and mortality and the emergence of new variants has reduced the effectiveness of current vaccines.Methods: Here, as a proof‐of‐concept, we engineered primary CD8 T cells to express SARS‐CoV‐2 Spike protein‐specific CARs, using the extracellular region of ACE2 and demonstrated their highly specific and potent cytotoxicity towards Spike‐expressing target cells. To improve on this concept as a potential therapeutic, we developed a bispecific T cell engager combining ACE2 with an anti‐CD3 scFv (ACE2‐Bite) to target infected cells and the virus.Results: As in CAR‐T cell approach, ACE2‐Bite endowed cytotoxic cells to selectively kill Spike‐expressing targets. Furthermore, ACE2‐Bite neutralized the pseudoviruses of SARS‐CoV, SARS‐CoV‐2 wild‐type, and variants including Delta and Omicron, as a decoy protein. Remarkably, ACE2‐Bite molecule showed a higher binding and neutralization affinity to Delta and Omicron variants compared to SARS‐CoV‐2 wild‐type Spike proteins.Conclusion: In conclusion, these results suggest the potential of this approach as a variant‐proof, therapeutic strategy for future SARS‐CoV‐2 variants, employing both humoral and cellular arms of the adaptive immune response.