There is a need for better classification and understanding of tumor-infiltrating lymphocytes (TILs). Here, we applied advanced functional genomics to interrogate 9,000 human tumors and multiple ...single-cell sequencing sets using benchmarked T cell states, comprehensive T cell differentiation trajectories, human and mouse vaccine responses, and other human TILs. Compared with other T cell states, enrichment of T memory/resident memory programs was observed across solid tumors. Trajectory analysis of single-cell melanoma CD8+ TILs also identified a high fraction of memory/resident memory-scoring TILs in anti-PD-1 responders, which expanded post therapy. In contrast, TILs scoring highly for early T cell activation, but not exhaustion, associated with non-response. Late/persistent, but not early activation signatures, prognosticate melanoma survival, and co-express with dendritic cell and IFN-γ response programs. These data identify an activation-like state associated to poor response and suggest successful memory conversion, above resuscitation of exhaustion, is an under-appreciated aspect of successful anti-tumoral immunity.
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•Improved global TIL classification methods are required to deconvolve cell states•αPD-1 non-responder TILs and dysfunctional TILs score for T activation, not exhaustion•αPD-1 response and patient survival associates with late T cell memory/TRM scoring•Persistent programs co-express with DC maturation and IFN-γ response programs
Jaiswal et al. highlight the need for improved tumor-infiltrating lymphocyte (TIL) classification by showing that current transcriptome assignments may misclassify early activated/effector TILs as exhausted. The study surveys 9,000 solid tumors, multiple single-cell RNA sequencing sets, mouse and human models, and scoring methods to reclassify TILs and associate melanoma survival to T cell memory/resident memory.
BackgroundImmune checkpoint inhibitors (ICIs) are limited by the high incidence of immune-related adverse events (irAEs) occurring in up to 40% of solid tumor patients on anti-PD-1 monotherapy 1 2 ...and 72% in anti-CTLA-4/anti-PD-1 combination.3 4 These toxicities can cause treatment cessation, hospitalization and even death.5–7 IrAEs are variable in severity, timing, onset, and remain poorly understood. Amongst the different toxicities, skin irAEs are most frequent, occur the earliest, and are correlated with a positive prognosis.4 8 However, there is a lack of preclinical models to study checkpoint toxicity. We evaluated a murine model of allergic contact dermatitis (contact hypersensitivity to 2,4-dinitrofluorobenzene) that is mediated by CD8+ T cells to gain a mechanistic understanding of skin checkpoint toxicity.MethodsC57BL/6 mice (n = 5 per group) were sensitized epicutaneously on shaved flank with hapten 0.5% DNFB on day -5 and elicited on their ears with DNFB on day 0. Starting four weeks later, mice were treated with either anti-programmed cell death protein (PD-1) or isotype. At the time of the first recall challenge only, mice were given either anti-PD-1 or isotype. Mice received subsequent rechallenges with DNFB to the ears and ear swelling was measured at various time points. Mice were depleted of circulating or skin CD8+ T cells by anti-CD8 mAbs from day 29 onwards, and maintained weekly, as in this model CD8+ T cells are the main hapten responder population. Samples were collected for histochemistry and analyzed by flow cytometry.ResultsOur data indicate that despite the depletion of circulating T cells, anti-PD-1 recipients mount a higher initial recall response to contact agents. Higher ear swelling was observed with increased inflammation in these mice. Our data suggest anti-PD-1 can liberate local T cell responses in the absence of a contribution from blood, and may offer a model to test therapeutic interventions to alleviate peripheral immune toxicities.ConclusionsOur results suggest that this murine model of contact hypersensitivity represents a potential model for skin immune checkpoint toxicities. This model of locally-mediated inflammatory recall may advance the goal of uncoupling toxicity from efficacy in patients with immune-related adverse events.Ethics ApprovalThe animal study was approved by Weill Cornell Medicine’s IACUC; approval number D16-00186.ReferencesNaidoo J, Page DB, Li BT, et al. Toxicities of the anti-PD-1 and anti-PD-L1 immune checkpoint antibodies. Ann Oncol 2015;26(12):2375–91. doi: 10.1093/annonc/mdv383.Belum VR, Benhuri B, Postow MA, et al. Characterisation and management of dermatologic adverse events to agents targeting the PD-1 receptor. Eur J Cancer 2016;60:12–25. doi: 10.1016/j.ejca.2016.02.010.Postow MA, Sidlow R, Hellmann MD. Immune-Related Adverse Events Associated with Immune Checkpoint Blockade. N Engl J Med 2018;378(2):158–168. doi: 10.1056/NEJMra1703481.Martins F, Sofiya L, Sykiotis GP, et al. Adverse effects of immune-checkpoint inhibitors: epidemiology, management and surveillance. Nat Rev Clin Oncol 2019;16(9):563–580. doi: 10.1038/s41571-019-0218-0.Puzanov I, Diab A, Abdallah K, et al. Managing toxicities associated with immune checkpoint inhibitors: consensus recommendations from the society for immunotherapy of cancer (SITC) Toxicity Management Working Group. J Immunother Cancer 2017;5(1):95. doi: 10.1186/s40425-017-0300-z.Wills B, Brahmer JR, Naidoo J. Treatment of complications from immune checkpoint inhibition in patients with lung cancer. Curr Treat Options Oncol 2018;19(9):46. doi: 10.1007/s11864-018-0562-9.Michot JM, Bigenwald C, Champiat S, et al. Immune-related adverse events with immune checkpoint blockade: a comprehensive review. Eur J Cancer 2016;54:139–148. doi: 10.1016/j.ejca.2015.11.016.Phillips GS, Wu J, Hellmann MD, et al. Treatment outcomes of immune-related cutaneous adverse events. J Clin Oncol 2019:JCO1802141. doi: 10.1200/JCO.18.02141.
Study Type – Therapy (case series)
Level of Evidence 4
OBJECTIVE
• To evaluate the safety and efficacy of ultrasonography (US)‐guided renal access in percutaneous nephrolithotomy (PCNL), as compared ...with conventional fluoroscopy‐guided renal access in a prospective randomized trial.
PATIENTS AND METHODS
• From January 2008 to October 2009, 224 patients with renal calculi undergoing PCNL were randomized into two groups.
• Group 1 (112 patients) underwent PCNL using only fluoroscopy‐guided renal access; while in group 2 (112 patients), US guidance for puncture was used in addition to fluoroscopy.
• The inclusion criteria were: normal renal functions, American Society of Anesthesiology scores 1 or 2, absence of congenital abnormalities, aged 15–70 years, and anticipated single‐tract procedure. The patients in both groups were matched for age, sex, and stone characteristics.
• The Student t‐test was used for statistical analysis with an allowable error of 5%.
RESULTS
• The mean time to successful puncture was 3.2 min and 1.8 min in group 1 and group 2, respectively (P < 0.01).
• The mean duration of radiation exposure to successful puncture was 28.6 s in group 1 and 14.4 s in group 2 (P < 0.01).
• The mean numbers of attempts for successful puncture in the desired calyx was 3.3 in group 1 as compared with 1.5 in group 2 (P < 0.01).
• The meantime taken for tract formation in group 1 was 7.4 min with radiation exposure of 82 s, while in group 2 it took 4.8 min with radiation exposure of 58 s (P < 0.01).
• Successful access was achieved in all patients. All patients were stone‐free at the end of the operation. The hospital stay (2–3 days) was same in both groups. There was no incidence of significant bleeding requiring transfusion during or after surgery. All the patients were followed‐up for a ≥6 months.
CONCLUSION
• US‐guided puncture in PCNL helps in increasing accuracy of puncture and decreasing radiation exposure for the surgical team and the patients.
Alopecia areata (AA) is among the most prevalent autoimmune diseases, but the development of innovative therapeutic strategies has lagged due to an incomplete understanding of the immunological ...underpinnings of disease. Here, we performed single-cell RNA sequencing (scRNAseq) of skin-infiltrating immune cells from the graft-induced C3H/HeJ mouse model of AA, coupled with antibody-based depletion to interrogate the functional role of specific cell types in AA in vivo. Since AA is predominantly T cell-mediated, we focused on dissecting lymphocyte function in AA. Both our scRNAseq and functional studies established CD8+ T cells as the primary disease-driving cell type in AA. Only the depletion of CD8+ T cells, but not CD4+ T cells, NK, B, or γδ T cells, was sufficient to prevent and reverse AA. Selective depletion of regulatory T cells (T
) showed that T
are protective against AA in C3H/HeJ mice, suggesting that failure of T
-mediated immunosuppression is not a major disease mechanism in AA. Focused analyses of CD8+ T cells revealed five subsets, whose heterogeneity is defined by an "effectorness gradient" of interrelated transcriptional states that culminate in increased effector function and tissue residency. scRNAseq of human AA skin showed that CD8+ T cells in human AA follow a similar trajectory, underscoring that shared mechanisms drive disease in both murine and human AA. Our study represents a comprehensive, systematic interrogation of lymphocyte heterogeneity in AA and uncovers a novel framework for AA-associated CD8+ T cells with implications for the design of future therapeutics.
Antigen-specific CD8
T cell accumulation in tumors is a prerequisite for effective immunotherapy, and yet the mechanisms of lymphocyte transit are not well defined. Here we show that tumor-associated ...lymphatic vessels control T cell exit from tumors via the chemokine CXCL12, and intratumoral antigen encounter tunes CXCR4 expression by effector CD8
T cells. Only high-affinity antigen downregulates CXCR4 and upregulates the CXCL12 decoy receptor, ACKR3, thereby reducing CXCL12 sensitivity and promoting T cell retention. A diverse repertoire of functional tumor-specific CD8
T cells, therefore, exit the tumor, which limits the pool of CD8
T cells available to exert tumor control. CXCR4 inhibition or loss of lymphatic-specific CXCL12 boosts T cell retention and enhances tumor control. These data indicate that strategies to limit T cell egress might be an approach to boost the quantity and quality of intratumoral T cells and thereby response to immunotherapy.
In this paper, we propose a model for estimating short-channel effects (SCEs) in the shell-doped double-gate junctionless (JL) MOSFET. The main emphasis of this paper is to estimate SCEs by ...effectively capturing source/drain (S/D) extensions beyond the gate edges for different values of undoped core thickness (<inline-formula> <tex-math notation="LaTeX">{T}_{\textsf {core}} </tex-math></inline-formula>), shell doping (<inline-formula> <tex-math notation="LaTeX">{N}_{\textsf {d}} </tex-math></inline-formula>), gate length (<inline-formula> <tex-math notation="LaTeX">{L}_{\textsf {g}} </tex-math></inline-formula>), and gate (<inline-formula> <tex-math notation="LaTeX">{V}_{\textsf {gs}} </tex-math></inline-formula>) and drain (<inline-formula> <tex-math notation="LaTeX">{V}_{\textsf {ds}} </tex-math></inline-formula>) biases in subthreshold regime. The threshold voltage (<inline-formula> <tex-math notation="LaTeX">{V}_{\textsf {th}} </tex-math></inline-formula>), drain-induced barrier lowering and subthreshold swing (<inline-formula> <tex-math notation="LaTeX">{S} </tex-math></inline-formula>), extracted from transfer characteristics, are in good agreement with the simulation results. Results highlight the utility of shell doping and core thickness to provide an additional degree of freedom to control SCEs. The proposed model can be useful in estimating SCEs and optimizing core-shell JL architecture for low-power applications.
In this paper, we investigate the impact of gate-source/drain underlap on short-channel behavior of junctionless (JL) transistor through a quasi-analytical model and 2-D numerical simulations. The ...proposed five-region model for potential is developed for the symmetric mode operation of double-gate (DG) JL MOSFET in the subthreshold regime, to predict and minimize short-channel effects (SCEs) using the optimum design of underlap regions. The five-region model can be transformed into four or three regions to incorporate the possible dependence of biases, doping, and gate workfunction with the underlap length. The parameters indicating SCEs can also be extracted using an analytical approximation for subthreshold drain current. The results from the proposed model are in reasonable agreement with simulation data. Analyses show that underlap length is a critical parameter to restrict the lateral extension of depletion region into the ungated portion. With increasing the underlap length, SCEs improve prominently for moderate doping concentration (<inline-formula> <tex-math notation="LaTeX">10^{18} </tex-math></inline-formula> cm<inline-formula> <tex-math notation="LaTeX">^{-3} </tex-math></inline-formula>). The improvement in SCEs ceases once the device achieves the maximum lateral extension of the depletion region. This paper provides the physical insights into the optimal use of underlap region in nanoscale JL devices for suppressing SCEs.
This paper proposes a semianalytical model to estimate short-channel effects for independent gate operation in double-gate (DG) junctionless (JL) MOSFET incorporating gate-to-source/drain underlap, ...through the solution of Poisson's equations in the subthreshold regime. The model also accounts for the asymmetry in device operation through variation in gate oxide thicknesses, gate work functions, and underlap lengths. Subthreshold drain current, threshold voltage, and subthreshold swing, evaluated from the channel potential, show reasonable agreement with simulation data. Results suggest the use of negative back gate bias and longer underlap length to reduce off-current. This paper highlights the role of doping, underlap length, and back gate bias in tuning the threshold voltage. This model serves as a generic formulation (within limits) with different asymmetries to estimate, design, and optimize self-aligned DG JL transistors for subthreshold logic applications.
NF-E2-related factor 2 (Nrf2) regulates expression and coordinated induction of a battery of chemoprotective genes in response to oxidative and electrophilic stress. This leads to protection against ...oxidative stress and neoplastic diseases. Nuclear import and export of Nrf2 play a significant role in control of nuclear levels of Nrf2 and thus the expression of Nrf2 down-stream genes. Tyrosine kinase Fyn phosphorylates tyrosine 568 of Nrf2 that leads to the nuclear export of Nrf2. In this study, we investigated the upstream factor(s) in regulation of Fyn and Fyn-mediated nuclear export of Nrf2. The investigations shed light on a novel mechanism of Nrf2 regulation in response to oxidative stress. We demonstrate that GSK-3β acts upstream of Fyn kinase in control of nuclear export of Nrf2. Chemical and short interfering RNA-mediated inhibition of GSK-3β led to nuclear accumulation of Nrf2 and transcriptional activation of the Nrf2 downstream gene nqo1. Chemical and short interfering RNA inhibition of GSK-3β and Fyn individually and in combination revealed that both kinases follow the same pathway to regulate nuclear export of Nrf2. We further demonstrate that hydrogen peroxide phosphorylates tyrosine 216 of GSK-3β. This leads to activation of GSK-3β. The activated GSK-3β phosphorylates Fyn at threonine residue(s). Phosphorylated Fyn accumulates in the nucleus and phosphorylates Nrf2 at tyrosine 568. This leads to nuclear export, ubiquitination, and degradation of Nrf2.
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