Therapeutic potential of CRISPR/CAS9 genome modification in T cell-based immunotherapy of cancer.

Today, genome editing technologies like zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR) are being used in clinical trials and the treatment of diseases like acquired immunodeficiency syndrome (AIDS) and cancer. CRISPR stands out as one of the most advanced tools for genome editing due to its simplicity and cost-effectiveness. It can selectively modify specific locations in the genome, offering new possibilities for treating human diseases. The CRISPR system uses ribonucleic acid-deoxyribonucleic acid (RNA-DNA) recognition to combat infections, regulate gene expression, and treat cancer. Chimeric antigen receptor (CAR) T-cell therapy, which uses T lymphocytes to eliminate cancer cells, can be improved by combining it with CRISPR technology. However, there are challenges in using CAR-T cells, including a lack of quantity and quality, exhaustion, neurotoxicity, cytokine release syndrome (CRS), B cell aplasia, tumor lysis syndrome, and anaphylaxis. Preclinical studies on CRISPR-edited CAR-T cells show promising results and targeting detrimental regulatory genes can enhance cancer treatment in the future.

Hepatoprotective effect of amifostine and WR-1065 on acetaminophen-induced liver toxicity on Wistar rats.

PURPOSE: The most important problem with acetaminophen is its hepatotoxicity. N-acetylcysteine (NAC) is used to treat the hepatotoxicity of acetaminophen. Due to the structural similarities of this compound with amifostine, we decided to test the effect of this substance and its metabolite, WR-1065, on the hepatotoxicity of acetaminophen. METHODS: The single-dose method contained 1. Control; 2. Acetaminophen (1 g/kg, gavage); 3-5. Acetaminophen + amifostine (100, 200, 400 mg/kg, i.p.); 6-8. Acetaminophen + WR-1065 (50, 100, 200 mg/kg, i.p.); and 9. Acetaminophen + NAC (100, 200 mg/kg, i.p.). The multiple-dose method included the same groups: amifostine (50, 100, 200 mg/kg), WR-1065 (25, 50, 100 mg/kg), and NAC (100 mg/kg). Then, animals were sacrificed, and blood samples were collected for measuring ALT, AST, ALP, and T-Bil, liver tissue for histopathological examination, MDA, and GSH amounts. RESULTS: Acetaminophen increased the levels of MDA, T-Bil, ALT, AST, and ALP, decreased GSH levels, and augmented necrosis, neutrophils, lymphocytes, and macrophages in the port space in single-dose and multiple-dose studies. Amifostine and WR-1065 significantly reduced the levels of MDA, T-Bil, ALT, AST, ALP, increased GSH content, and ameliorated histopathological alterations in a single-dose and multiple-dose method compared to the acetaminophen group. Moreover, NAC caused a significant decrease in the levels of MDA, T-Bil, ALT, AST, and ALP, and reduced GSH amounts in single-dose and multiple-dose studies. CONCLUSION: Amifostine and WR-1065 as antioxidant and hepatoprotective compounds are effective in reducing acetaminophen-induced hepatotoxicity with a similar effect to NAC and can be administered as an adjunct in the treatment of acetaminophen overdose.

Autonomous swab robot for naso- and oropharyngeal COVID-19 screening.

The COVID-19 outbreak has triggered a global health and economic crisis, necessitating widespread testing to control viral spread amidst rising cases and fatalities. The recommended testing method, a combined naso- and oropharyngeal swab, poses risks and demands limited protective gear. In response to the COVID-19 pandemic, we developed and tested the first autonomous swab robot station for Naso- and Oropharyngeal Coronavirus Screening (SR-NOCS). A force-sensitive robot running under a Cartesian impedance controller is employed to drive the swab to the sampling area. This groundbreaking device underwent two clinical studies-one conducted during the initial pandemic lockdown in Europe (early 2021) and the other, more recently, in a public place after the pandemic had subsided earlier in the year 2023. In total, 52 patients suspected of COVID-19 infection were included in these clinical studies. The results revealed a complete positive correlation between autonomous and manual sampling. The test subjects exhibited a high acceptance rate, all expressing a willingness to undergo future tests with SR-NOCS. Based on our findings, such systems could enhance testing capabilities, potentially conducting up to 300 tests per robot per day with consistent precision. The tests can be carried out with minimal supervision, reducing infection risks and effectively safeguarding patients and healthcare workers.

Model-Based Shared Control of a Hybrid FES-Exoskeleton: An Application in Participant-Specific Robotic Rehabilitation.

Hybrid exoskeleton, comprising an exoskeleton interfaced with functional electrical stimulation (FES) technique, is conceptualized to complement the weakness of each other in automated neuro-rehabilitation of sensory-motor deficits. The externally actuating exoskeleton cannot directly influence neurophysiology of the patients, while FES is difficult to use in functional or goal-oriented tasks. The latter challenge is largely inherited from the fact that the dynamics of the muscular response to FES is complex, and it is highly user- and state-dependent. Due to the retardation of the muscular contraction response to the FES profile, furthermore, a commonly used model-free control scheme, such as PID control, suffers performance. The challenge in FES control is exacerbated especially in the presence of the actuation redundancy between the volitional activity of the user, powered exoskeleton, and FES-induced muscle contractions. This study therefore presents trajectory tracking performance of the hybrid exoskeleton in a novel model-based hybrid exoskeleton scheme which entices user-specific FES model-predictive control.

Comparing self-medication and medical supervised treatment before hospitalization among patients with Covid-19: A retrospective case-control study.

BACKGROUND: During the Covid-19 pandemic, there has been a notable increase in self-medication with antibiotics or other medications due to impaired access to healthcare services. This kind of self-treatment, without comprehending the condition and its related risks, can result in misdiagnosis, overdosing and delaying in acquiring professional medical attention, or may even cause antimicrobial resistance. Additionally, reports have suggested that medical practitioners have prescribed medications inappropriately to patients with Covid-19. To investigate this further, this study compared the medications used by patients with Covid-19 prior to hospitalization with or without a medical recommendation. METHODS: Data was extracted a mass survey of patients with of Covid-19 in Mashhad, and the patients were divided into two main groups: those who received medication with guidance from a medical professional (treatment group) and those who self-administered medications without professional oversight (self-treatment group). Statistical analysis was then conducted using SPSS version 26, the Chi-square, and multiple logistic regression test. RESULTS: This study examined 3266 patients, with 1466 included in the analysis. Results showed that men (9.5 %), those living in rural areas (21 %), and those with no academic degree (37.5%) had a higher likelihood to self-medicating. Antibiotics were the most frequently used medications prior to hospitalization (9.5%). Comparing the two groups revealed that three drug categories- antibiotics, antivirals and other medications (medicines that are not in the other 4 main categories)- were utilized more often in the treatment group than in the self-treatment group, with a p-value of < 0.05. The only medical condition that had a significant difference between the two groups was diabetes, with 34.1 % in the self-treatment group versus 24.5 % in the treatment group (P < 0.05). CONCLUSIONS: The Covid-19 pandemic has caused a surge in the inappropriate use of certain medications through self-medicating. This poses a serious risk to the health of patients, highlighting the need for not only adjusting guidelines but also raising awareness and enforcing compliance to prevent unnecessary use of drugs.

Investigating the Anti-tumor and Apoptosis-inducing Effects of Coumarin Derivatives as Potent 15-Lipoxygenase Inhibitors on PC-3 Prostate Cancer Cells.

INTRODUCTION: Prostate cancer is the second most prevalent cancer among men. Despite different treatments, including surgery, chemotherapy, radiation therapy, hormone therapy and immunotherapy for this disease, patients ultimately progress to advanced states. Thus, there is a need for new treatment options targeting cell growth and apoptosis to better control the proliferation and metastasis of these cells. There are many reports indicating overexpression of the 15-lipoxygenase-1 (15-LOX-1) enzyme in prostate tumors. Studies have also shown that inhibition of this enzyme prevents the progression of prostate cancer. OBJECTIVE: This study was conducted to assess the anti-cancer properties of some coumarin derivatives as possible 15- LOX-1 inhibitors, on PC-3 prostate cancer cells. METHODS: In this study, the activity of 15-LOX-1 was evaluated in PC-3 cells by a spectrophotometric assay. In addition, due to high similarity between the 15-LOX-1 and soybean 15-lipoxygenase (SLO) (L1; EC 1, 13, 11, 12) active sites, the soybean SLO was used to investigate inhibitory effects of synthetic coumarin compounds 8- isopentenyloxycoumarin (8-IC), 8-isopentenyloxy-3-carboxycoumarin (8-ICC), 8-geranyloxycoumarin (8-GC), 8- geranyloxy-3-carboxycoumarin (8-GCC), and 8-farnesyloxy-3-carboxycoumarin (8-FCC) on this enzyme. Moreover, the cytotoxic and anticancer effects of the coumarin compounds were examined on PC-3 (Prostate Cancer) and HDF-1 (Human Dermal Fibroblast) cells by alamarBlue assay. Finally, apoptosis-inducing effects of all synthetic compounds were determined by flow cytometry. RESULTS: The IC50 values obtained by the alamarBlue test revealed that 8-IC, 8-GC and 8-GCC had cytotoxic effects on PC-3 cells. Treating both PC-3 and HDF-1 cells with 8-ICC and 8-FCC did not significantly reduce cell number. Furthermore, the IC50 values of 8-IC on HDF-1 cells showed cytotoxic effects, while treating these cells with 8-GC and 8- GCC did not show any significant cytotoxicity on these normal human fibroblasts. Assessing the ability of 4-MMPB (as a specific inhibitor of 15-LOX-1), 8-GC, and 8-GCC compounds to inhibit SLO revealed that these compounds exerted strong 15-LOX-1 inhibitory activity, while 8-IC and 8-FCC had a weak inhibitory effect and also 8-ICC showed no inhibitory effect on SLO enzyme. In addition, flow cytometric analysis by FITC (fluorescein isothiocyanate)- annexin V and propidium iodide showed that treatment with IC50 values of 8-GC and 8-GCC induced apoptosis in 35.2% and 30.8% of PC-3 cells, respectively. CONCLUSION: Thus, 8-GC and 8-GCC can be introduced as effective anticancer agents with apoptosis-inducing properties. Furthermore, our results suggest that the cytotoxic effects of these compounds might be related to the inhibition of 15-LOX-1 enzyme in PC-3 cells. On the other hand, the cytotoxic effects of 8-IC might be due to the inhibition of other signaling pathways in PC-3 cells. However, further in vivo experiments are required to determine the exact mechanisms involved in the anticancer effects of these coumarin compounds.

A Comparative Study on Visual Detection of Mycobacterium tuberculosis by Closed Tube Loop-Mediated Isothermal Amplification: Shedding Light on the Use of Eriochrome Black T.

Loop-mediated isothermal amplification is a promising candidate for the rapid detection of Mycobacterium tuberculosis. However, the high potential for carry-over contamination is the main obstacle to its routine use. Here, a closed tube LAMP was intended for the visual detection of Mtb to compare turbidimetric and two more favorable colorimetric methods using calcein and hydroxy naphthol blue (HNB). Additionally, a less studied dye (i.e., eriochrome black T (EBT)) was optimized in detail in the reaction for the first time. Mtb purified DNA and 30 clinical specimens were used to respectively determine the analytical and diagnostic sensitivities of each method. The turbidimetric method resulted in the best analytical sensitivity (100 fg DNA/reaction), diagnostic sensitivity and specificity (100%), and time-to-positivity of the test (15 min). However, this method is highly prone to subjective error in reading the results. Moreover, HNB-, calcein-, and EBT-LAMP could respectively detect 100 fg, 1 pg, and 1 pg DNA/reaction (the analytical sensitivities) in 30, 15, and 30 min, while the diagnostic sensitivity and specificity were respectively 93.3% and 100% for them all. Interestingly, EBT-LAMP showed the lowest potential for subjective error in reading the results. This report helps judiciously choose the most appropriate visual method, taking a step forward toward the field applicability of LAMP for the detection of Mtb, particularly in resource-limited settings.

Effect of biosynthesized selenium nanoparticles using Nepeta extract against multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii.

The problems of drug resistance in bacteria have become one of the daily challenges of the clinical treatment of patients, which inevitably forces us to use agents other than common antibiotics. Among these, we can take help from different properties and applications of nanoparticles (NPs). In this work, we evaluate the antibacterial activity of biosynthesized selenium nanoparticles (SeNPs) against standard strains of multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii. The production of biosynthesized SeNPs was proved by ultraviolet-visible, Fourier transform infrared, X-ray diffractometer, Field Emission Scanning Electron Microscopy, Dynamic light scattering, and Zeta potential methods. The cytotoxicity effect of SeNPs was investigated by MTT assay. Disk diffusion agar (DDA) and minimum inhibitory concentration (MIC) tests were performed on the mentioned bacteria using different classes of standard antibiotics and SeNPs separately. The impact of SeNPs combined with the desired antibiotics for better treatment of these infections was evaluated by checkerboard assay to determine the synergism effect. After the confirmation results based on the biosynthesis of SeNPs, both standard bacterial strains were susceptible to SeNPs and had a zone of inhibition using the DDA test. Also, the results of MICs showed that biosynthesized SeNPs in lower concentrations than antibiotics cause no growth of bacteria. On the other hand, according to the checkerboard assay, SeNPs had a synergistic effect with conventional antibiotics. The antibacterial sensitivity tests demonstrated the inhibition of bacterial growth in the presence of lower concentrations of SeNPs than common antibiotics. This property can be exerted in future applications to solve the drug resistance obstacle of microorganisms in bacterial diseases.

Tactile Robotic Telemedicine for Safe Remote Diagnostics in Times of Corona: System Design, Feasibility and Usability Study.

The current crisis surrounding the COVID-19 pandemic demonstrates the amount of responsibility and the workload on our healthcare system and, above all, on the medical staff around the world. In this work, we propose a promising approach to overcome this problem using robot-assisted telediagnostics, which allows medical experts to examine patients from distance. The designed telediagnostic system consists of two robotic arms. Each robot is located at the doctor and patient sites. Such a system enables the doctor to have a direct conversation via telepresence and to examine patients through robot-assisted inspection (guided tactile and audiovisual contact). The proposed bilateral teleoperation system is redundant in terms of teleoperation control algorithms and visual feedback. Specifically, we implemented two main control modes: joint-based and displacement-based teleoperation. The joint-based mode was implemented due to its high transparency and ease of mapping between Leader and Follower whereas the displacement-based is highly flexible in terms of relative pose mapping and null-space control. Tracking tests between Leader and Follower were conducted on our system using both wired and wireless connections. Moreover, our system was tested by seven medical doctors in two experiments. User studies demonstrated the system's usability and it was successfully validated by the medical experts.

LZTR1 molecular genetic overlap with clinical implications for Noonan syndrome and schwannomatosis.

BACKGROUND: Noonan syndrome (NS) is a genetic disorder characterized by developmental delays, typical facial gestalt and cardiovascular defects. LZTR1 variants have been recently described in patients with NS and schwannomatosis, but the association, inheritance pattern and management strategy has not been fully elucidated. Here, we review the contribution of LZTR1 in NS and describe a patient with a novel, likely pathogenic variant in LZTR1. CASE PRESENTATION: A female patient was diagnosed with clinical NS at 8 months of age. She presented in adulthood when a brain and spine MRI identified plexiform neurofibromas; however, she did not meet the clinical criteria for Neurofibromatosis type 1. No pathogenic variants were identified through molecular genetic analysis of NF1, SPRED1 and a multigene NS panel. Whole exome sequencing at age 23 identified a novel de novo likely pathogenic heterozygous variant in the LZTR1 gene denoted as c.743G>A (p.Gly248Glu). Serial MRIs have shown stable imaging findings and the patient is being followed clinically by cardiology, neurology and medical genetics. CONCLUSIONS: We identified a novel mutation in the LZTR1 gene, not previously reported in association with NS. This report provides additional evidence to support for the assessment of schwannomatosis in patients with LZTR1-NS and may have overlap with Neurofibromatosis type 1.

Munich Institute of Robotics and Machine Intelligence (MIRMI) Technical University of Munich. / Mensch-Roboter-Interaktion: vernetzte, lernfähige Maschinen in der Medizin.

The application of robotic and intelligent technologies in healthcare is dramatically increasing. The next generation of lightweight and tactile robots have provided a great opportunity to be used for a wide range of applications from medical examination, diagnosis, therapeutic procedures to rehabilitation and assistive robotics. They can potentially outperform current medical procedures by exploiting the com- plementary strengths of humans and computer-based technologies. In this study, the importance of human- robot interaction is discussed and technological re- quirements and challenges in making human-centered robot platforms for medical applications is addressed.

Guillain-Barré syndrome as an early manifestation of angioimmunoblastic T-cell lymphoma.

Guillain-Barré syndrome (GBS) is a rare condition caused by autoimmune damage of peripheral nerves. We describe a case where a man in his 80s presented with subacute, progressive fatigue and weakness. He had received an outpatient work-up for possible haematological malignancy, but eventually presented to the emergency department for worsening weakness. A physical exam and cerebrospinal fluid analysis suggested a diagnosis of GBS. Subsequently, a pathological diagnosis of angioimmunoblastic T-cell lymphoma was made. The patient underwent intravenous immunoglobulin treatment for GBS and was started on cyclophosphamide, doxorubicin, vincristine and prednisone therapy. Prior research has suggested that incident malignancy may be associated with GBS, which may be caused by a paraneoplastic-type phenomenon, malignancy-associated immune dysregulation or an autoimmune reaction triggered by a common exposure. Clinicians should be aware of the possible association between these two conditions and should remain open minded to the possibility of non-infectious triggers for GBS.

8-Geranyloxycarbostyril as a potent 15-LOX-1 inhibitor showed great anti-tumor effects against prostate cancer.

Carbostyrils are quinolone derivatives, with possible growth inhibition properties on cancer cells. Unlike many tumors, 15-Lipoxygenase-1 (15-LOX-1) is highly expressed in prostate cancer (PCa) cells and has oncogenic properties. Here, with the hypothesis that 6-, 7- and 8-geranyloxycarbostyril (GQ) have inhibitory properties on 15-LOX-1, their effects were assessed on PCa cells. Their cytotoxic effects were evaluated by MTT assay and mechanism of cell death was investigated using annexin V/PI staining. Finally, the anti-tumor properties of 8-GQ were assessed in immunocompromised C57BL/6 mice bearing human PCa cells. Accordingly, these compounds could effectively inhibit 15-LOX activity in PCa cells. MTT and flow cytometry tests confirmed their toxic effects on PCa cells, with no significant toxicity on normal cells, and apoptosis was the main mechanism of cell death. In vivo results indicated that use of 8-GQ at 50 mg/kg had stronger anti-tumor effects than 5 mg/kg cisplatin, with fewer side effects on normal tissues. Therefore, 8-GQ can be introduced as a potential drug candidate with 15-LOX-1 inhibitory potency, which can be effective in treatment of prostate cancer, and should be considered for further drug screening investigations.

O-prenylated carbostyrils as a novel class of 15-lipoxygenase inhibitors: Synthesis, characterization, and inhibitory assessment.

Catalyzed peroxidation of unsaturated lipid in animals and plants intimately is linked to the activity of 15-Lipoxygenase enzymes. Lipoxygenases (LOXs) are well known to play an important role in many acute and chronic syndromes such as inflammation, asthma, cancer, and allergy. In this study, a series of mono prenyloxycarbostyrils were synthesized and evaluated as potential inhibitors of soybean 15-Lipoxygenase (SLO) and their inhibitory potencies were compared to mono prenyloxycoumarins which had been reported in the previous works. The synthetic compounds inhibit lipoxygenase enzyme by competitive mechanism like the prenyloxy coumarins. The results showed that position and length of the prenyl moiety play the important role in lipoxygenase inhibitory activity. Among all of the synthetic compounds (coumarin and carbostyril derivatives), 5-farnesyloxycoumarin and 8-farnesyloxycarbostyril demonstrated the best inhibitory activity by IC50  values of 1.1 µM and 0.53 µM, respectively.

Investigating the effects of two novel 4-MMPB analogs as potent lipoxygenase inhibitors for prostate cancer treatment.

BACKGROUND: Lipoxygenases are one of the critical signaling mediators which can be targeted for human prostate cancer (PC) therapy. In this study, 4-methyl-2-(4-methylpiperazinyl)pyrimido[4,5-b]benzothiazine (4-MMPB) and its two analogs, 4-propyl-2-(4-methylpiperazinyl)pyrimido[4,5-b]benzothiazine (4-PMPB) and 4-ethyl-2-(4-methylpiperazinyl)pyrimido[4,5-b]benzothiazine (4-EMPB), were proposed to have anti-tumor properties in prostate cancer. METHODS: After synthesizing the compounds, cytotoxic effects of 4-MMPB and its two analogs against PC-3 cancerous and HDF normal cells were investigated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and then mechanism of cell death was assessed by flow cytometry. Finally, the anti-tumor effects of the mentioned compounds were investigated in an immunocompromised C57BL/6 mouse model. RESULTS: 4-PMPB and 4-EMPB had similar anti-cancer effects on PC-3 cells as compared with 4-MMPB, while they were not effective on normal cells. Moreover, apoptosis and ferroptosis were the main mechanisms of induced cell death in these cancerous cells. Furthermore, in vivo results indicated that both analogs had similar anti-cancer effects as 4-MMPB, leading to delayed tumor growth without any noticeable side effects in weight loss and histological investigations. CONCLUSION: Thus, our results suggest that specific targeting of lipoxygenases via 4-MMPB analogs can be considered as a treatment of choice for PC therapy, although it requires further investigations.

Design, synthesis, and structure-activity relationship study of O-prenylated 3-acetylcoumarins as potent inhibitors of soybean 15-lipoxygenase.

In this work, the design, synthesis, and structure-activity relationships of a novel array of geranyloxy and farnesyloxy 3-acetylcoumarins were reported as potent soybean 15-lipoxygenase inhibitors. Among the prepared coumarins, 7-farnesyloxy-3-acetylcoumarin (12b) was found to be the most potent inhibitor by IC50  = 0.68 µM while O-geranyl substituents at positions 5 and 6 of 3-acetylcoumarin (10a and 11a) were not inhibitors. Using docking studies, the binding affinity and the preferred pose of synthetic compounds were considered. It was found that lipoxygenase inhibitory activity and prenyl length chain were directly related. The hydrophobic cavity of the enzyme was more effectively occupied by the farnesyl moiety of the potent inhibitor 12b rather than other derivatives. Also, with this pose of farnesyl chain in 7-farnesyloxy-3-acetylcoumarins, the acetyl group could be directed to the hydrophilic pocket in the active site.

Allylphenols as a new class of human 15-lipoxygenase-1 inhibitors.

In this study, a series of mono- and diallylphenol derivative were designed, synthesized, and evaluated as potential human 15-lipoxygenase-1 (15-hLOX-1) inhibitors. Radical scavenging potency of the synthetic allylphenol derivatives was assessed and the results were in accordance with lipoxygenase (LOX) inhibition potency. It was found that the electronic natures of allyl moiety and para substituents play the main role in radical scavenging activity and subsequently LOX inhibition potency of the synthetic inhibitors. Among the synthetic compounds, 2,6-diallyl-4-(hexyloxy)phenol (42) and 2,6-diallyl-4-aminophenol (47) showed the best results for LOX inhibition (IC50 = 0.88 and 0.80 µM, respectively).

Design, synthesis and evaluation of PD-L1 peptide antagonists as new anticancer agents for immunotherapy.

Blocking the interaction of programmed cell death protein 1 (PD-1) and its ligand PD-L1 is known as a promising immunotherapy for treatment of a variety of tumors expressing PD-L1 on their cell surface. In the last decade, several antibodies against the PD-1/PD-L1 interaction have been approved, while there are few reports of small-molecule inhibitors against PD-1/PD-L1 axis. Due to many advantages of cancer treatment with small molecules over antibodies, we developed several peptidic PD-L1 antagonists using computational peptide design methods, and evaluated them both in vitro and in vivo. Importantly, among six peptides with best affinity to PD-L1, four peptides exhibited significant potency to block PD-1/PD-L1 axis at molecular level. Moreover, the PD-L1 expression in nine human colorectal cancer cell lines stimulated with interferon-γ was compared and LoVo cells with the highest expression were selected for further experiments. The peptides could also restore the function of activated Jurkat T cells, which had been suppressed by stimulated LoVo cells. A blockade assay in tumor-bearing mice experiments indicated that peptides HS5 and HS6 consisting of a d-amino acid in their structures, could also effectively reduce tumor growth in vivo, without induction of any observable liver or renal toxicity, tissue damages and loss of body weight. As new designed peptides showed no toxicity against murine colon cancer cells in vitro, the observed anti-tumor results in mice are most probably due to disrupting the PD-1/PD-L1 interaction. Thus, peptides described in this study can be considered as proper low molecular weight candidates for immunotherapy of cancer.