Magnetic Field Emulations of Small Inhibitor RNA: Effects on Implanted GL261 Tumors in C57BL/6 Immune Competent Mice

EMulate Therapeutics has developed a system for emulating the effects of solvated molecules via their magnetic field recordings. Recordings of magnetic field emissions of select small inhibitor RNAs (siRNAs;murine targeting CTLA-4 and murine targeting PD-1) were tested on C57Bl/6 mice implanted subcutaneously with the GL261 murine tumor cell line. A signal composed of concatenated recordings of siRNA molecules targeting the murine CTLA-4 and PD-1 receptors (labeled A2) was used in immune competent C57Bl/6 mice. The mice were flank implanted with the murine glioblastoma cell line GL261. Mice were exposed to the signal continuously (24 hours a day) until tumor volumes reached the designated volume limit. Tumors were excised and analyzed via PAGE/Western blot for the expression of CTLA-4, PD-1, Ki67, Caspase 3, CD4 and CD8. Terminal blood draws were used for CBCs. We report the down regulation of the checkpoint inhibitors CTLA-4 in the exposed mice. Significant tumor volume reduction was observed in mice exposed to the siRNA signal compared to control mice;no adverse events were recorded. Cell blood counts (CBC) and protein expression patterns were observed to correlate with the expected function of protein expression inhibition of the targets.

Genomic correlates of the response to first-line PD-1 blockade plus chemotherapy in patients with advanced non-small-cell lung cancer

Background:Programmed death 1(PD-1)blockade plus chemotherapy has become the new first-line standard of care for patients with advanced non-small-cell lung cancer(NSCLC).Yet not all NSCLC patients benefit from this regimen.This study aimed to investigate the predictors of PD-1 blockade plus chemotherapy in untreated advanced NSCLC.Methods:We integrated clinical,genomic,and survival data from 287 patients with untreated advanced NSCLC who were enrolled in one of five registered phase 3 trials and received PD-1 blockade plus chemotherapy or chemotherapy alone.We randomly assigned these patients into a discovery cohort(n=125),a validation cohort(n=82),and a control cohort(n=80).The candidate genes that could predict the response to PD-1 blockade plus chemotherapy were identified using data from the discovery cohort and their predictive values were then evaluated in the three cohorts.Immune deconvolution was conducted using transcriptome data of 1014 NSCLC patients from The Cancer Genome Atlas dataset.Results:A genomic variation signature,in which one or more of the 15 candidate genes were altered,was correlated with significantly inferior response rates and survival outcomes in patients treated with first-line PD-1 blockade plus chemotherapy in both discovery and validation cohorts.Its predictive value held in multivariate analyses when adjusted for baseline parameters,programmed cell death ligand 1(PD-L1)expression level,and tumor mutation burden.Moreover,applying both the 15-gene panel and PD-L1 expression level produced better performance than either alone in predicting benefit from this treatment combination.Immune landscape analyses revealed that tumors with one or more variation in the 15-gene panel were associated with few immune infiltrates,indicating an immune-desert tumor microenvironment.Conclusion:These findings indicate that a 15-gene panel can serve as a negative prediction biomarker for first-line PD-1 blockade plus chemotherapy in patients with advanced NSCLC.

Aerosolized immunotherapeutic nanoparticle inhalation potentiates PD-L1 blockade for locally advanced lung cancer

Despite therapeutic advancements,the prognosis of locally advanced non-small cell lung cancer(LANSCLC),which has invaded multiple lobes or the other lung and intrapulmonary lymph nodes,remains poor.The emergence of immunotherapy with immune checkpoint blockade(ICB)is transforming cancer treatment.However,only a fraction of lung cancer patients benefit from ICB.Significant clinical evidence suggests that the proinflammatory tumor microenvironment(TME)and programmed death-ligand 1(PD-L1)expression correlate positively with response to the PD-1/PD-L1 blockade.We report here a liposomal nanoparticle loaded with cyclic dinucleotide and aerosolized(AeroNP-CDN)for inhalation delivery to deep-seated lung tumors and target CDN to activate stimulators of interferon(IFN)genes in macrophages and dendritic cells(DCs).Using a mouse model that recapitulates the clinical LANSCLC,we show that AeroNP-CDN efficiently mitigates the immunosuppressive TME by reprogramming tumor-associated macrophage from the M2 to M1 phenotype,activating DCs for effective tumor antigen presentation and increasing tumor-infiltrating CD8+T cells for adaptive anticancer immunity.Intriguingly,activation of interferons by AeroNP-CDN also led to increased PD-L1 expression in lung tumors,which,however,set a stage for response to anti-PD-L1 treatment.Indeed,anti-PD-L1 antibody-mediated blockade of IFNs-induced immune inhibitory PD-1/PD-L1 signaling further prolonged the survival of the LANSCLC-bearing mice.Importantly,AeroNP-CDN alone or combination immunotherapy was safe without local or systemic immunotoxicity.In conclusion,this study demonstrates a potential nano-immunotherapy strategy for LANSCLC,and mechanistic insights into the evolution of adaptive immune resistance provide a rational combination immunotherapy to overcome it.

PD-1／PD-L1通路相关的肾癌免疫治疗研究进展

肾细胞癌（RCC）是泌尿外科常见肿瘤之一，占成人全部恶性肿瘤的2％～3％，接受根治性肾切除的肾癌患者有20％～40％会逐渐出现复发或转移。对于转移性的晚期肾细胞癌患者，结合靶向治疗的综合治疗是主要的治疗方式。肾癌的免疫治疗研究曾一度进展缓慢，近年来随着程序性死亡受体1（Programmed cell death 1 receptor，PD-1）／（Programmed cell death 1 ligand 1，PD—L1）通路的研究进展，

A novel PD-L1 targeting peptide self-assembled nanofibers for sensitive tumor imaging and photothermal immunotherapy in vivo

Programmed death 1(PD-1)and its ligand PD-L1 are two typical immune checkpoints.Antibody-based immune checkpoint blockade(ICB)strategy targeting PD-1/PD-L1 achieved a significant therapeutic effect on cancer.However,due to the impenetrability of antibody drugs and the occurrence of immune-related adverse events,only a minority of patients benefit from this treatment.Peptides multimerization has been widely proved to be an effective method to improve receptor binding affinity through a multivalent synergistic effect.In this study,we report a novel peptide-aggregation-induced emission(AIE)hybrid supramolecular TAP,which can self-assemble into nanofibers through non-covalent interactions such as hydrogen bonds,with a specific nanomolar affinity to PD-L1 in vivo and in vitro.Combined with near-infrared agents,it can be used for tumor imaging and photothermal therapy,which enables photothermal ablation of cancer cells for generating tumor-associated antigen(TAA)and triggering a series of immunological events.Collectively,our work suggests that synthetic self-assembled peptide nanofibers can be developed as attractive platforms for active photothermal immunotherapies against cancer.

Assays for predicting and monitoring responses to lung cancer immunotherapy

Immunotherapy has become a key strategy for cancer treatment, and two immune checkpoints, namely, programmed cell death 1 （PD-1） and its ligand （PD-L1）, have recently emerged as important targets. The interaction blockade of PD-1 and PD-L1 demonstrated promising activity and antitumor efficacy in early phase clinical trials for advanced solid tumors such as non-small cell lung cancer （NSCLC）. Many cell types in multiple tissues express PD-L1 as well as several tumor types, thereby suggesting that the ligand may play important roles in inhibiting immune responses throughout the body. Therefore, PD-L1 is a critical immunomodulating component within the lung microenvironment, but the correlation between PD-L1 expression and prognosis is controversial. More evidence is required to support the use of PD-L1 as a potential predictive biomarker. Clinical trials have measured PD-L1 in tumor tissues by immunohistochemistry （IHC） with different antibodies, but the assessment of PD-L1 is not yet standardized. Some commercial antibodies lack specificity and their reproducibility has not been fully evaluated. Further studies are required to clarify the optimal IHC assay as well as to predict and monitor the immune responses of the PD-I/PD-L1 pathway.

Advances in immunotherapy for treatment of lung cancer

Different approaches for treating lung cancer have been developed over time, including chemotherapy, radiotherapy and targeted therapies against activating mutations. Lately, better understanding of the role of the immunological system in tumor control has opened multiple doors to implement different strategies to enhance immune response against cancer cells. It is known that tumor cells elude immune response by several mechanisms. The development of monoclonal antibodies against the checkpoint inhibitor programmed cell death protein 1 （PD-1） and its ligand （PD-L1）, on T cells, has led to high activity in cancer patients with long lasting responses. Nivolumab, an anti PD-1 inhibitor, has been recently approved for the treatment of squamous cell lung cancer patients, given the survival advantage demonstrated in a phase III trial. Pembrolizumab~ another anti PD-1 antibod）5 has received FDA breakthrough therapy designation for treatment of non-small cell lung cancer （NSCLC）, supported by data from a phase I trial. Clinical trials with anti PD-1/PD-L1 antibodies in NSCLC have demonstrated very good tolerability and activity, with response rates around 20% and a median duration of response of 18 months.

Fueling the engine and releasing the break: combinational therapy of cancer vaccines and immune checkpoint inhibitors

Immune checkpoint inhibitors are increasingly drawing much attention in the therapeutic development for cancer treatment. However, many cancer patients do not respond to treatments with immune checkpoint inhibitors, partly because of the lack of tumor-infiltrating effector T cells. Cancer vaccines may prime patients for treatments with immune checkpoint inhibitors by inducing effector T-ceU infiltration into the tumors and immune checkpoint signals. The combination of cancer vaccine and an immune checkpoint inhibitor may function synergistically to induce more effective antitumor immune responses, and clinical trials to test the combination are currently ongoing.

Camrelizumab(SHR-1210)treatment for recurrent hepatocellular carcinoma after liver transplant:A report of two cases

Immune checkpoint inhibitors are generally contraindicated for post-transplant patients.However,we report two patients with metastatic hepatocellular carcinoma(HCC)treated with camrelizumab(SHR-1210),an anti-programmed cell death 1(PD-1)agent,after liver transplant.Before undergoing immunotherapy,both patients underwent liver allograft biopsy and obtained negative PD-L1 expression in tumor and liver graft specimens by immunohistochemistry.Then,camrelizumab(200 mg)was administered once every 3 weeks.During immunotherapy,the targeted therapy was continued,and the immunosuppression regimen was adjusted to a low-dose level.No graft rejection or other severe adverse reactions were observed.The disease remained stable(SD,mRECIST)for 3 months in one patient and 10 months in the other.Therefore,camrelizumab may have safety and potential benefits in advanced HCC after liver transplant.

Non-small cell lung cancer in China

In China,lung cancer is a primary cancer type with high incidence and mortality.Risk factors for lung cancer include tobacco use,family history,radiation exposure,and the presence of chronic lung diseases.Most early-stage non-small cell lung cancer(NSCLC)patients miss the optimal timing for treatment due to the lack of clinical presentations.Population-based nationwide screening programs are of significant help in increasing the early detection and survival rates of NSCLC in China.The understanding of molecular carcinogenesis and the identification of oncogenic drivers dramatically facilitate the development of targeted therapy for NSCLC,thus prolonging survival in patients with positive drivers.In the exploration of immune escape mechanisms,programmed cell death protein 1(PD-1)/programmed death-ligand 1(PD-L1)inhibitor monotherapy and PD-1/PD-L1 inhibitor plus chemotherapy have become a standard of care for advanced NSCLC in China.In the Chinese Society of Clinical Oncology’s guidelines for NSCLC,maintenance immunotherapy is recommended for locally advanced NSCLC after chemoradiotherapy.Adjuvant immunotherapy and neoadjuvant chemoimmunotherapy will be approved for resectable NSCLC.In this review,we summarized recent advances in NSCLC in China in terms of epidemiology,biology,molecular pathology,pathogenesis,screening,diagnosis,targeted therapy,and immunotherapy。

Mechanisms of immune checkpoint inhibitormediated liver injury

The immune checkpoints,cytotoxic T-lymphocyte-associated antigen 4(CTLA-4) and programmed cell death protein-1/ligand-1(PD-1/PD-L1) are vital contributors to immune resulation and tolerance.Recently immune checkpoint inhibitors(ICIs) have revolutionized cancer therapy;however,they come with the cost of immune related adverse events involving multiple organs such as the liver.Due to its constant expo sure to foreign antigens,the liver has evolved a high capacity for immune tolerance,therefore,blockade of the immune checkpoints can result in aberrant immune activation affecting the liver in up to 20% of patients depending on the agent(s) used and underlying factors.This type of hepatotoxicity is termed immune mediated liver injury from checkpoint inhibitors(ILICI) and is more common when CTLA4 and PD-1/PDL1 are used in combination.The underlying mechanisms of this unique type of hepatotoxicity are not fully understood;however,the contribution of CD8^(+) cytotoxic T lymphocytes,various CD4^(+) T cells populations,cytokines,and the secondary activation of the innate immune system leading to liver injury have all been suggested.This review summarizes our current understanding of the underlying mechanisms of liver injury in immunotherapy using animal models of ILICI and available patient data from clinical studies.

Immune checkpoint:The novel target for antitumor therapy

Inhibitory checkpoint molecules include programmed cell death-1(PD-1),programmed cell death ligand-1(PD-L1),cytotoxic T lymphocyte antigen-4(CTLA-4),human endogenous retrovirus-H Long terminal repeat-associating 2(HHLA2),B7 homolog 4 protein(B7-H4),T cell membrane protein-3(TIM-3)and Lymphocyte-activation gene 3(LAG-3),which are up-regulated during tumorigenesis.These pathways are essential to down-regulate the immune system by blocking the activation of T cells.In recent years,immune checkpoint blockers(ICBs)against PD-1,PD-L1,CTLA-4 or TIM-3 has made remarkable progress in the clinical application,revolutionizing the treatment of malignant tumors and improving patients’overall survival.However,the efficacy of ICBs in some patients does not seem to be good enough,and more immune-related adverse events(irAEs)will inevitably occur.Therefore,biomarkers research provides practical guidance for clinicians to identify patients who are most likely to benefit from or exhibit resistance to particular types of immune checkpoint therapy.There are two points in general.On the one hand,given the spatial and temporal differential expression of immune checkpoint molecules during immunosuppression process,it is essential to understand their mechanisms to design the most effective individualized therapy.On the other hand,due to the lack of potent immune checkpoints,it is necessary to combine them with novel biomarkers(such as exosomes and ctDNA)and other anticancer modalities(such as chemotherapy and radiotherapy).

Immune checkpoints and immunotherapy for colorectal cancer

Colorectal cancer(CRC)remains one of the major causes of death worldwide,despite steady improvement in early detection and overall survival over the past decade.Current treatment paradigms,with chemotherapy and biologics,appear to have reached their maximum benefit.Immunotherapy,especially with checkpoint inhibitors,has shown considerable clinical benefit in various cancers,including mismatch-repair-deficient CRC.This has led to the planning and initiation of several clinical trials evaluating novel immunotherapy agents—as single agents,combinations and in conjunction with chemotherapy—in patients with CRC.This article reviews biological and preclinical data for checkpoint inhibitors and discusses various immunotherapy trials in CRC,as well as current efforts in CRC immunotherapy.