Stem Cell Marker Expression in Early Stage Colorectal Cancer is Associated with Recurrent Intestinal Neoplasia.

BACKGROUND: Colorectal cancer (CRC) ranks second in cancer deaths worldwide and presents multiple management challenges, one of which is identifying high risk stage II disease that may benefit from adjuvant therapy. Molecular biomarkers, such as ones that identify stem cell activity, could better stratify high-risk cohorts for additional treatment. METHODS: To identify possible biomarkers of high-risk disease in early-stage CRC, a discovery set (n = 66) of advanced-stage tumors were immunostained with antibodies to stemness proteins (CD166, CD44, CD26, and LGR5) and then digitally analyzed. Using a second validation cohort (n = 54) of primary CRC tumors, we analyzed protein and gene expression of CD166 across disease stages, and extended our analyses to CD166-associated genes (LGR5, ASCL2, BMI1, POSTN, and VIM) by qRT-PCR. RESULTS: Stage III and metastatic CRC tumors highly expressed stem cell-associated proteins, CD166, CD44, and LGR5. When evaluated across stages, CD166 protein expression was elevated in advanced-stage compared to early-stage tumors. Notably, a small subset of stage I and II cancers harbored elevated CD166 protein expression, which correlated with development of recurrent cancer or adenomatous polyps. Gene expression analyses of CD166-associated molecules revealed elevated ASCL2 in primary tumors from patients who recurred. CONCLUSIONS: We identified a protein signature prognostic of aggressive disease in early stage CRC. Stem cell-associated protein and gene expression identified a subset of early-stage tumors associated with cancer recurrence and/or subsequent adenoma formation. Signatures for stemness offer promising fingerprints for stratifying early-stage patients at high risk of recurrence.

Defining a stem cell hierarchy in the intestine: markers, caveats and controversies.

The past decade has appreciated rapid advance in identifying the once elusive intestinal stem cell (ISC) populations that fuel the continual renewal of the epithelial layer. This advance was largely driven by identification of novel stem cell marker genes, revealing the existence of quiescent, slowly- and active-cycling ISC populations. However, a critical barrier for translating this knowledge to human health and disease remains elucidating the functional interplay between diverse stem cell populations. Currently, the precise hierarchical and regulatory relationships between these ISC populations are under intense scrutiny. The classical theory of a linear hierarchy, where quiescent and slowly-cycling stem cells self-renew but replenish an active-cycling population, is well established in other rapidly renewing tissues such as the haematopoietic system. Efforts to definitively establish a similar stem cell hierarchy within the intestinal epithelium have yielded conflicting results, been difficult to interpret, and suggest non-conventional alternatives to a linear hierarchy. While these new and potentially paradigm-shifting discoveries are intriguing, the field will require development of a number of critical tools, including highly specific stem cell marker genes along with more rigorous experimental methodologies, to delineate the complex cellular relationships within this dynamic organ system.

Cost-Effectiveness of Continuous Support From a Layperson During a Woman's First Two Births.

OBJECTIVE: To evaluate the cost-effectiveness and health outcomes related to continuous support from a layperson during a woman's first two births in a theoretical population. DESIGN: Cost-effectiveness analysis. PARTICIPANTS: A theoretical cohort of 1.2 million women based on an approximation of annual low-risk, nulliparous, term, singleton births in the United States with the assumption that these women have second births. This reflects the average number of births per woman in the United States. METHODS: We designed a cost-effectiveness model to compare outcomes in women with continuous support from relatives, friends, or community members with minimal to no training (excluding trained doulas) during labor and birth compared with outcomes for women with no continuous support. Outcomes included mode of birth, uterine rupture, hysterectomy, maternal death, cost, and quality-adjusted life years (QALYs). We derived probabilities from the literature and set a cost-effectiveness threshold at $100,000/QALY. RESULTS: In this theoretical model, continuous support by a layperson during the first birth resulted in fewer cesarean births, decreased costs, and increased QALYs for the first and subsequent births. Women with support from laypersons had 71,090 fewer cesarean births, 35 fewer uterine ruptures, 9 fewer hysterectomies, and 16 fewer maternal deaths, which saved $364 million with 2,673 increased QALYs. Sensitivity analyses showed that continuous support in the first birth was cost-effective even when varying the estimate of lost wages of the support person up to $708. CONCLUSION: Continuous labor support from a layperson leads to fewer cesarean births, improved outcomes, decreased costs, and increased QALYs. This highlights the need to increase women's access to continuous layperson support during labor and birth uninhibited by financial and institutional barriers.

The Cost-Effectiveness of Professional Doula Care for a Woman's First Two Births: A Decision Analysis Model.

INTRODUCTION: Multiple studies have demonstrated the benefits of intrapartum doula care, including lower risk for cesarean birth and shortened labor time for nulliparous women. However, analyses investigating the cost-effectiveness of doula care are limited. This study evaluated the potential cost-effectiveness of professional doula support during a woman's first birth in a theoretical population of US women, with all women having a second birth without doula care. METHODS: A cost-effectiveness model was designed to compare outcomes in women with a professional doula versus no doula labor support. A theoretical cohort of 1.6 million women, the approximate number of annual low-risk, nulliparous, term, singleton births in the United States, was used. Outcomes included mode of birth, maternal death, uterine rupture, cesarean hysterectomy, costs, and quality-adjusted life years (QALYs). Probability estimates used in the model were derived from the literature, and a cost-effectiveness threshold was set at $100,000 per QALY. Sensitivity analyses were used to investigate the robustness of the results. RESULTS: In this theoretical model, professional doula care during the first birth resulted in fewer cesarean births and improved QALYs. Additionally, doula support resulted in 202,538 fewer cesarean births, 46 fewer maternal deaths secondary to fewer cesarean births, 99 fewer uterine ruptures, and 26 fewer hysterectomies, with an additional cost of $185 million and 7617 increased QALYs for the first and subsequent births. Sensitivity analyses demonstrated a professional doula was potentially cost-saving up to $884 and cost-effective up to $1360 per doula. DISCUSSION: Professional doula care during a woman's first birth may lead to improved outcomes and increased QALYs during her first and second births. Given the limitations of this analysis, the cost-effectiveness estimate is likely conservative, further supporting broader integration of professional doulas into the US maternity care system and highlighting the need for higher doula care reimbursement.

Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations.

BACKGROUND & AIMS: Continual renewal of the intestinal epithelium is dependent on active- and slow-cycling stem cells that are confined to the crypt base. Tight regulation of these stem cell populations maintains homeostasis by balancing proliferation and differentiation to support critical intestinal functions. The hierarchical relation of discrete stem cell populations in homeostasis or during regenerative epithelial repair remains controversial. Although recent studies have supported a model for the active-cycling leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5)+ intestinal stem cell (ISC) functioning upstream of the slow-cycling B lymphoma Mo-MLV insertion region 1 homolog (Bmi1)-expressing cell, other studies have reported the opposite relation. Tools that facilitate simultaneous analyses of these populations are required to evaluate their coordinated function. METHODS: We used novel monoclonal antibodies (mAbs) raised against murine intestinal epithelial cells in conjunction with ISC-green fluorescent protein (GFP) reporter mice to analyze relations between ISC populations by microscopy. Ex vivo 3-dimensional cultures, flow cytometry, and quantitative reverse-transcription polymerase chain reaction analyses were performed. RESULTS: Two novel mAbs recognized distinct subpopulations of the intestinal epithelium and when used in combination permitted isolation of discrete Lgr5GFP and Bmi1GFP-enriched populations with stem activity. Growth from singly isolated Lgr5GFP ISCs gave rise to small spheroids. Spheroids did not express Lgr5GFP and instead up-regulated Bmi1GFP expression. Conversely, Bmi1-derived spheroids initiated Lgr5GFP expression as crypt domains were established. CONCLUSIONS: These data showed the functional utility of murine mAbs in the isolation and investigation of Lgr5GFP and Bmi1GFP ISC-enriched populations. Ex vivo analyses showed hierarchical plasticity between different ISC-expressing states; specifically Lgr5GFP ISCs gave rise to Bmi1GFP cells, and vice versa. These data highlight the impact of temporal and physiological context on unappreciated interactions between Lgr5GFP and Bmi1GFP cells during crypt formation.

Cell Adhesion Molecule CD166/ALCAM Functions Within the Crypt to Orchestrate Murine Intestinal Stem Cell Homeostasis.

BACKGROUND & AIMS: Intestinal epithelial homeostasis is maintained by active-cycling and slow-cycling stem cells confined within an instructive crypt-based niche. Exquisite regulating of these stem cell populations along the proliferation-to-differentiation axis maintains a homeostatic balance to prevent hyperproliferation and cancer. Although recent studies focus on how secreted ligands from mesenchymal and epithelial populations regulate intestinal stem cells (ISCs), it remains unclear what role cell adhesion plays in shaping the regulatory niche. Previously we have shown that the cell adhesion molecule and cancer stem cell marker, CD166/ALCAM (activated leukocyte cell adhesion molecule), is highly expressed by both active-cycling Lgr5+ ISCs and adjacent Paneth cells within the crypt base, supporting the hypothesis that CD166 functions to mediate ISC maintenance and signal coordination. METHODS: Here we tested this hypothesis by analyzing a CD166-/- mouse combined with immunohistochemical, flow cytometry, gene expression, and enteroid culture. RESULTS: We found that animals lacking CD166 expression harbored fewer active-cycling Lgr5+ ISCs. Homeostasis was maintained by expansion of the transit-amplifying compartment and not by slow-cycling Bmi1+ ISC stimulation. Loss of active-cycling ISCs was coupled with deregulated Paneth cell homeostasis, manifested as increased numbers of immature Paneth progenitors due to decreased terminal differentiation, linked to defective Wnt signaling. CD166-/- Paneth cells expressed reduced Wnt3 ligand expression and depleted nuclear ß-catenin. CONCLUSIONS: These data support a function for CD166 as an important cell adhesion molecule that shapes the signaling microenvironment by mediating ISC-niche cell interactions. Furthermore, loss of CD166 expression results in decreased ISC and Paneth cell homeostasis and an altered Wnt microenvironment.