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Glycans are carbohydrate molecules appended to proteins and lipids on the surface of all living cells. Glycans play key roles in a wide array of biological processes, and structural changes in cell-surface glycosylation patterns have been connected to pathogenesis of several diseases. In particular, cancer cells frequently upregulate expression of glycans that bind to inhibitory receptors (lectins) on immune cells. These glycosylated antigens systematically inhibit immune activity and protect cancer cells from immune surveillance. Understanding how cancer cells generate these glycan ligands can thus lead to identification of novel druggable targets for therapeutic intervention. However, glycan ligand biosynthesis is subject to extremely complex genetic regulation, making it difficult to identify the key genes involved in production of immune-regulatory glycan antigens. In a recent publication, we described a CRISPR/Cas9 screening approach to identify genes that drive synthesis of ligands for glycan-binding immune receptors. Here, we outline a detailed, step-by-step protocol for completing this type of genome-wide screen. Our protocol produces a genome-wide atlas of all genes whose expression is required for cell-surface binding of a recombinant immune lectin. This dataset can be used both to identify novel ligands for immune lectins and annotate regulatory genes that drive changes in cancer-associated glycosylation. Our protocol serves as a general resource for researchers interested in the detailed study of cancer glyco-immunology. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Generation of a genome-wide CRISPR library using lentiviral transduction Support Protocol: Generation of dCas9KRAB-expressing K-562 cells Basic Protocol 2: Staining of genome-wide CRISPR libraries with Siglec-Fc reagents and fluorescence-activated cell sorting Basic Protocol 3: Library amplification and sequencing Basic Protocol 4: Data analysis and hit identification., (© 2023 Wiley Periodicals LLC.)

Magrolimab (Hu5F9-G4) is a first-in-class anti-CD47 IgG4 monoclonal antibody, with potential applications in several malignancies including myelodysplastic syndrome. CD47 blockade in malignancy has been shown to promote antitumour effects. However, the ubiquity of CD47 on red blood cells can result in interference in pretransfusion immunohaematology investigations and hinder timely provision of red blood cell units, with potential to mask clinically significant alloantibodies. We reviewed the literature for pretransfusion interference seen with magrolimab and methods to circumvent potential issues, and sought to provide clinical and laboratory recommendations for safe local transfusion practices. These recommendations are based on expert opinion and available literature, including the Victorian Senior Transfusion Scientist working group and professional societies and organisations (Australian & New Zealand Society of Blood Transfusion and Lifeblood representatives), to establish consensus recommendations. Interference in the ABO group and antibody screen can occur, and baseline immunohaematology testing prior to magrolimab therapy is critical. Antibody screening using an antihuman globulin reagent that does not detect human IgG4 subclass may distinguish magrolimab interference from an underlying alloantibody in patient plasma. Clear and consistent protocols for laboratories and close communication with clinicians are paramount to facilitate timely and safe transfusion support for patients receiving magrolimab therapy. As local transfusion laboratories gain experience with magrolimab, this will assist in our understanding and comfort in managing these patients., (© 2022 The Authors. Internal Medicine Journal published by John Wiley & Sons Australia, Ltd on behalf of Royal Australasian College of Physicians.)

Competing Interests: Nicolo Matteo Luca BattistiConsulting or Advisory Role: Pfizer, Sanofi, Abbott NutritionSpeakers' Bureau: Pfizer, AbbVie, SanofiTravel, Accommodations, Expenses: Pfizer, Genomic Health, Lilly Mark LawlerHonoraria: Pfizer, Roche, Bayer Health, NovartisTravel, Accommodations, Expenses: Pfizer, Roche Katie RizviHonoraria: Roche Robert GreeneConsulting or Advisory Role: GlaxoSmithKline Kathy OliverConsulting or Advisory Role: Bristol Myers Squibb (Inst), Seattle Genetics (Inst), Novartis Italy (Inst), Sanofi/Regeneron (Inst), Novocure (Inst), Eisai (Inst)Other Relationship: Bristol Myers Squibb (Inst), Novocure (Inst), Photonamic (Inst), Bayer (Inst), Northwest Biotherapeutics (Inst), Novartis (Inst), Elekta (Inst), Medac (Inst), GW Pharmaceuticals (Inst), Pfizer (Inst), Karyopharm Therapeutics (Inst), Apogenix (Inst)No other potential conflicts of interest were reported.

Background: Early postoperative enteral nutrition with immune-enhancing supplements has helped to restore immune function and reduce infectious complications in patients with cancer undergoing major gastrointestinal operations. The aim of this study was to evaluate the effectiveness of similar supplements (containing arginine and arginine plus omega-3 fatty acids) given preoperatively for 1 week before cancer surgery., Methods: In this randomized, double-blinded study, patients scheduled to undergo elective resection of upper gastrointestinal tumors were given one of three different oral liquid supplemental diets (control, arginine, arginine plus omega-3 fatty acids) to be taken each day for 7 days before surgery. Blood samples were obtained upon enrollment, on the morning of surgery, and on postoperative day 1 for analysis of immunologic function., Results: Mean serum ornithine (a metabolite of arginine) levels were significantly higher compared with controls, but no significant increase in mean serum arginine levels was noted on the morning of surgery for those patients who received arginine as part of the supplement. In conjunction with these findings, there were no differences among groups in mean lymphocyte mitogenesis, mean peripheral blood mononuclear cell production of cytokines, or clinical outcomes., Conclusions: Use of oral liquid supplements in this fashion did not improve lymphocyte proliferation or monocyte functions in patients with cancer undergoing major surgery.

Malnutrition, a common problem in cancer patients, adversely affects survival and quality of life. It results from several factors that alter nutritional intake and cause massive metabolic disturbances. Anticancer therapies may compound the malnutrition. Optimal nutrition improves therapeutic modalities and the clinical course and outcome. Oral nutrition should be used whenever possible; in patients unable to ingest adequate amounts orally, enteral and parenteral feedings are safe and effective.

No clear role has yet been defined for the role of total parenteral nutrition in the management of the cancer patient, and this subject remains an area of controversy. Peri-operative nutritional support, if given for at least 10 days, may reduce morbidity and mortality in some patients, although this subgroup has not been fully characterized. There is no evidence that parenteral nutritional support benefits patients undergoing chemotherapy or radiotherapy in terms of an increased tumor response rate or prolongation of survival.

Symptom distress, mental health status, enforced social dependency and health perceptions were measured in two groups of cancer patients, one receiving home care services (n = 49) and the other receiving no such services (n = 11). Data were obtained at hospital discharge and 3 months later. Patients receiving home care demonstrated statistically significant improvement on mental health and social dependency; patients not receiving home care did not improve on any variable. After controlling for baseline scores, the home care group had significantly higher mental health status at the second interview than the no home care group.

We performed an economic analysis of the care provided to patients undergoing major abdominal surgical procedures to determine the effect of postoperative infection on hospital resource use. Patients' clinical and demographic characteristics and their use of medical care services were determined from a review of hospital bills and medical records. Hospital charges were obtained from the hospital billing system and costs were determined by use of Medicare cost-charge ratios obtained from the hospital's Medicare Cost Report. The care of patients with postoperative infections was significantly more expensive than that of uninfected patients (multivariate analysis indicated that a surgical infection added $12,542 to the cost of patient care). Patients with postoperative fever but without documented infection were also more expensive to care for than afebrile, uninfected patients (fever added $9145 to the cost of care). Increased costs for infected patients were found among microbiology tests, radiology services, pharmaceutical costs, and room costs. For these patients, we found that use of departmental cost-charge ratios, instead of hospital-wide cost-charge ratios, had no substantial impact on comparison of the cost of care for infected and uninfected patients.

Cancer patients are often at high risk for perioperative complications because of preexisting conditions, the magnitude of surgery, and the use of aggressive multimodality treatment. It is essential to identify risk factors preoperatively, correct any deficits, and monitor organ dysfunction. During the perioperative period prophylaxis and surveillance for cardiopulmonary, hematologic, and septic complications should minimize morbidity and mortality. Finally, nutritional support should be given to malnourished patients undergoing extensive operative procedures.

Cancer patients have the highest incidence of protein-calorie malnutrition seen in hospitalized patients, with significant malnutrition occurring in more than 30% of cancer patients undergoing major upper gastrointestinal procedures. Clinically significant malnutrition occurs as a result of diminished nutrient intake, increased nutrient losses, and tumor-induced derangements in host metabolism. In the absence of adequate exogenous nutrients, the body utilizes endogenous substrates to satisfy the ongoing requirements of both host and tumor for energy and protein. In those patients with malignant obstruction of the gastrointestinal tract, the tumor itself may induce diminished nutrient intake. Present day treatment modalities including gastrointestinal resection, chemotherapy, and radiotherapy compound these metabolic derangements, further increasing the risk of postoperative morbidity and death. The presence of malnutrition in cancer patients has prognostic importance. In a review of more than 3000 cancer patients, DeWys and colleagues identified significantly improved survival in those patients without weight loss compared with those had lost 6% of their body weight (Am J Med 69:491-497, 1980). Other investigators have noted increased postoperative morbidity and mortality associated with malnutrition. Early hypotheses suggested that reversal of weight loss would improve survival. The development and refinements of enteral and parenteral nutrition have provided the opportunity for studying the relationship between nutritional supplementation and postoperative prognosis. Nutrition support is therefore often instituted to improve nutritional status and thereby reduce the risks of postoperative complications. This article addresses the beneficial role of preoperative nutrition therapy in cancer patients.

Over 1,500 patients at our institution have received intravenous hyperalimentation (IVH) as nutritional support for nutritional rehabilitation prior to and/or during oncologic therapy. Stimulation of tumor growth has not been identified, and septic and metabolic complications of this technique have been minimal. Nutritional repletion resulted in return of immunocompetence and was associated with a reduction in sepsis, proper wound healing, and an apparent increase in tumor response to chemotherapy. If these observations were related as cause and effect, then a method of restoring and maintaining adequate nutrition should be added to the armamentarium of the oncologist. The use of IVh allowed specific oncologic therapy to be administered to a group of malnourished patients who otherwise might not have been acceptable candidates for intensive antineoplastic therapy.

The nutritional and immunological status of 140 malnourished cancer patients who were to receive intravenous hyperalimentation (IVH) were evaluated prior to treatment with IVH and chemotherapy (CMX), surgery or radiation therapy (XRT). Subsequently, these indices were correlated with responses to treatment. Cell-mediated immunocompetent (CMI+) patients (41%) had lost an average weight of 10 +/- 2.2 lbs., while cell-mediated immunoincompetent (CMI-) patients (59%) had lost an average weight of 31 +/- 2.5 lbs. (p less that 0.05). Total lymphocyte count (TLC) averaged 1290 +/- 250 cells/mm3 in CMI+ patients and 900 +/- 140 cells/mm" in CMI- patients (p less than 0.05). Serum albumin concentration (SA) was 3.6 +/- 0.06 g/dl in CMI+ patients and 3.3 +/- 0.05 g/dl in CMI- patients (p less than 0.05). In the CMI+ group, 49% had a positive response to CMX, whereas only 27% of the CMI-group responded to CMX (p less than 0.01). In the surgery group, 88% of the CMI+ patients, but only 65% of CMI- patients, had uncomplicated postoperative recovery periods. Although nutritional repletion with IVH was undertaken in all patients, those patients with initial CMI- responses had greater body weight loss, lower SA and TLC and either a decreased response to CMX or an increase in postoperative morbidity and mortality when compared with patients who initially had CMI+ responses.

Effects of nutritional repletion with intravenous hyperalimentation (IVH) on sequential skin test reactivity were evaluated in 160 malnourished cancer patients undergoing chemotherapy (76 patients), surgery (49 patients), radiation therapy (20 patients) and supportive care (15 patients). In the chemotherapy group, 45 patients had negative reactions initially, and 25 patients (55%) had at least one skin test convert to positive in an average period of 19 days of IVH. In the surgery group, 23 patients (46%) were initially positive and remained positive, 13 patients (24%) converted from negative to positive, and 13 patients (30%) remained negative or converted to negative. Postoperative complications occurred in 25% of positive reactors, compared with 69% (p < 0.01) of negative reactors. In the radiation therapy group, the skin tests of six patients (30%) remained positive, three patients (15%) converted from negative to positive and the skin tests of nine patients (45%) remained negative. In the supportive care group, the skin tests of 73% of the patients either remained positive or converted to positive with IVH within an average period of 11 days of treatment. Nutritional therapy with IVH was associated with restored skin test reactivity in 51% of malnourished cancer patients undergoing oncologic therapy. Radiation therapy was generally immunosuppressive despite adequate nutritional repletion. In surgical patients, positive skin test reactivity correlated directly with a favorable response to operative therapy.

An elective cancer education course at a public university attracted twice as many women as men. Course information was disseminated to an average of eight people by each student. The course seemed reasonably effective in stimulating positive behavioral changes. According to self-report, dietary fat was reduced by 47 per cent of women and 49 per cent of men. Fiber intake was increased by 48 per cent of women and 36 per cent of men. Breast self-examination was initiated by 36 per cent of previous nonperformers: and testicular self-examination, by 23 per cent.

Malnutrition is a common problem in cancer patients. The most clear-cut cause is anorexia and a decrease in food intake. Other causes include alterations in carbohydrate, fat and protein metabolism which result in losses of energy and amino acids from the host. Understanding the pathophysiology of these changes will provide avenues to better nourish cancer patients and improve their quality of life.

In over 1000 cancer patients treated with intravenous hyperalimentation (IVH), tumor growth has not been identified and catheter-related sepsis has been minimal. Studies in rats demonstrated that the host benefits more than the tumor during nutritional repletion, and any stimulation of tumor growth in the rat-tumor model could be manipulated with DNA specific drugs to benefit the host. A study of 65 malnourished cancer patients undergoing oncologic therapy and treated with IVH indicated that much of the immune suppression in these patients was the result of malnutrition coincident with or secondary to oncologic treatment. Conclusions reached in this study were that nutritional repletion resulted in a return of skin test reactivity, proper wound healing in the surgical patient, and possibly an increase in response to chemotherapy. Certainly, the use of IVH allowed specific oncologic therapy to be administered to a group of malnourished patients who otherwise might not have been acceptable candidates for intensive antineoplastic therapy.

Future horizons in surgical oncology relate to advances in prevention, diagnosis, and treatment of the patient with cancer. New diagnostic modalities may make complete surgical staging unnecessary. Perioperative support of patients given multimodality treatment will become more critical as the limits of surgical resection are reached and combined treatment with radiation and chemotherapy become more frequent. The current treatment of cancer at several organ sites reflects the evolving approach to treat both local and systemic manifestations of the malignancy. Use of intraoperative radiation therapy with electron-beam and/or brachytherapy has been proved safe and future prospective trials will determine efficacy in local control of disease. Finally, proper reconstruction becomes critical as we attempt to rehabilitate cancer patients toward normal function.

Cancer patients have the highest prevalence of malnutrition of any group of hospitalized patients. The potential causes of this malnutrition are numerous, as elements of both starvation and stress are evident in the cancer-bearing host. The presence of the tumor alone may lead to reduced intake of nutrients and treatment modalities of surgery, chemotherapy, and radiation therapy further exacerbate nutritional deficits. It is clear that the tumor requires energy substrates to grow, and that these substrates are exacted from the host. Animal studies identify progressive nutritional depletion concomitant with increasing tumor growth during ingestion of a regular diet. This appears predominantly due to reduced dietary intake in addition to host metabolic alterations. In animal/tumor models deliberate dietary protein depletion results in severe host weight loss, but also causes diminished tumor growth rates. Dietary manipulation in these animal/tumor models have demonstrated methods of improving tumor response to chemotherapy by manipulation of tumor growth rates. In addition, drug-pharmacokinetics have been altered by dietary manipulation. However, data from animal/tumor models are not directly applicable to man since the tumor in animals usually results in the death of the host within six to eight weeks. Nevertheless, controlled laboratory studies in animals provide basic metabolic information which promotes understanding of host/tumor relationships in man. In cancer patients malnutrition has prognostic value, leads to a distortion of body composition with erosion of body protein and fat stores, and compromises the delivery of adequate therapy. There is no direct objective evidence of accelerated tumor growth in humans with cancer who receive nutritional support as part of their treatment regimen. The host benefits to the extent that body composition is at least maintained during the period of nutritional repletion. Thus, nutritional support provides support to the patient during periods of treatment and dietary deprivation. No improvement in the tumor's response to therapy, however, has been demonstrated by this approach.

Surgery, radiation, and chemotherapy can result in further deterioration in nutritional status in patients who are frequently malnourished as a consequence of their underlying malignancy. Many of the effects are transient, and patients who have a good performance status may exhibit minimal alterations in nutritional parameters during the course of treatment. However, antitumor treatment may enhance morbidity, and in some, it leads to mortality if patients are in poor nutritional status ab initio. Such malnourished patients must be recognized prior to initiation of treatment, and steps must be taken to maintain adequate nutrition. The known toxic effects of many cancer therapeutic regimens must be weighed against their potential benefits.